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Operation Manual

IceCube 14S

Operator Manual for Software Version 1.21b and up

SY-LAB Geräte GmbH., Tullnerbachstrasse 61-65, A 3011 Neupurkersdorf, Austria


Tel +43 2231 622 52 –0, FAX +43 2231 621 93
Internet: http://www.sylab.com/, E-mail: sales@sylab.com, ©2000-2006 SY-LAB Geraete GmbH.
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Contents

Contents ___________________________________________________________________________ 2
Warranty ___________________________________________________________________________ 5
Purpose ___________________________________________________________________________ 6
Specification _______________________________________________________________________ 6
Freezer ________________________________________________________________________ 6
Power Connection________________________________________________________________ 6
Normal Conditions of Use __________________________________________________________ 7
Transportation and Storage Conditions _______________________________________________ 7
Additional Specification ____________________________________________________________ 7
Safety Regulations and Symbols ____________________________________________________ 7
Safety Notes ____________________________________________________________________ 8
Periodic Inspections ______________________________________________________________ 8
Liquid Nitrogen Dewar and Tubing ___________________________________________________ 8
Safety _____________________________________________________________________________ 9
General Precautions ______________________________________________________ 9
Special Precaution (Liquid Nitrogen, Tubing etc.) _____________________________ 10
Precaution for Transportation ______________________________________________ 10
Packing Contents ________________________________________________________ 11
Fuses __________________________________________________________________ 12
Important Specific Notes __________________________________________________ 13
Installation and Check-out ___________________________________________________________ 14
Site Preparation, Space and Clearance ______________________________________________ 14
Environment ___________________________________________________________________ 14
Unpacking ______________________________________________________________ 15
IceCube Installation - Wiring and Hook-Up ___________________________________ 17
Variant A: _____________________________________________________________________ 17
Variant B: _____________________________________________________________________ 17
Monitor Assembly _______________________________________________________________ 19
Variant C: _____________________________________________________________________ 20
Variant D: _____________________________________________________________________ 21
Connecting the LIN Hose to Freezer and Supply Vessel ________________________ 22
Pt-100 Transport Protection _______________________________________________ 22
Description of the Instrument Components and Connections ___________________ 23
Turn ON/OFF and Check of the System______________________________________________ 24
Handling of Temperature Sensors __________________________________________ 25
Freezing Chamber _______________________________________________________ 27
Description ____________________________________________________________________ 27
Available Freezing Racks _________________________________________________ 28
Program Log-In ____________________________________________________________________ 33
Description of Program Tools and Buttons ___________________________________ 34
Menu Area ____________________________________________________________________ 36
Main Menu - File ______________________________________________________________ 36
Edit description _____________________________________________________________ 36

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Open historical data _________________________________________________________ 39


Process directory _________________________________________________________ 39
Open last run ____________________________________________________________ 39
Set source directory _______________________________________________________ 39
Print Diagram, Print Program & Events, Print both _______________________________ 39
Close window ____________________________________________________________ 39
List of Available Tools ____________________________________________________ 42
Show description, Show program, Display Total View ___________________________________ 42
Create / Edit freezing program _________________________________________________ 43
Set data directory ___________________________________________________________ 43
Set program directory________________________________________________________ 44
Exit ______________________________________________________________________ 45
Main Menu - Display ___________________________________________________________ 46
Total View_________________________________________________________________ 46
Tracking View ______________________________________________________________ 46
Settings __________________________________________________________________ 46
Time axis _______________________________________________________________ 46
Temperature axis _________________________________________________________ 47
Time elapsed / remaining __________________________________________________ 47
Actual program position ____________________________________________________ 47
Temperature value to track _________________________________________________ 47
LNi pressure_____________________________________________________________ 47
Main Menu - Options __________________________________________________________ 48
Description of the various conditions and alarms __________________________________ 49
Start Temperature reached condition _________________________________________ 49
LNi pressure Alarm (not implemented on 14S) __________________________________ 49
Allowed Temperature deviation ______________________________________________ 50
Temperature reading invalid ________________________________________________ 50
Critical power fail - awaiting user assistance ____________________________________ 50
Short time power fail ______________________________________________________ 50
Freezing program finished __________________________________________________ 51
Save Settings ______________________________________________________________ 51
User Administration _________________________________________________________ 52
Info for the Administrator ___________________________________________________ 54
Language _________________________________________________________________ 55
Main Menu - Version Info ________________________________________________________ 56
Main Menu - Maintenance ________________________________________________________ 56
Ice-Water Calibration _________________________________________________________ 56
Communication _____________________________________________________________ 57
Writing a Test Program ___________________________________________________ 58
Test program for finding the crystallization temperature of a sample ________________________ 58
Freezing program editor ______________________________________________________ 59
Edit control - Program _____________________________________________________ 59
Edit Freezing program points________________________________________________ 59
Axis standard _____________________________________________________________ 62
Mouse button _____________________________________________________________ 62
Active point size___________________________________________________________ 62
Moving Screens and Zoom Functions____________________________________________ 62
Running the Test Program ________________________________________________________ 63
Operation Panel ____________________________________________________________ 66
Seeding ________________________________________________________________ 66
Hold ___________________________________________________________________ 66
Next ___________________________________________________________________ 66
Load program/Stop _______________________________________________________ 66
Writing the final program - Optimization (as an example) ______________________________ 68
Special Function - Autoseeding ________________________________________________ 72
Setting the Autoseeding Level _______________________________________________ 72
Setting the Restart Level ___________________________________________________ 76

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Seeding ___________________________________________________________________________ 78
Auto(matic)Seeding ______________________________________________________ 78
Maintenance _______________________________________________________________________ 79
Leak Check __________________________________________________________________ 79
Cleaning of the Freezing Chamber and Instrument ____________________________ 79
Handling of Temperature Sensors ________________________________________________ 79
Cleaning ____________________________________________________________________ 79
Settings on the PC __________________________________________________________________ 80
Disabling of Start-Up Error Message ___________________________________________________ 80
Freezer Software (Re-) Installation ____________________________________________________ 83
Inserting Your Own LOGO ___________________________________________________________ 90
Troubleshooting ___________________________________________________________________ 91
Freezer and Accessories ____________________________________________________________ 96
Furniture _______________________________________________________________ 96
Spare Parts 14S ____________________________________________________________________ 97
Appendix _________________________________________________________________________ 98
Statement of Conformity (220 - 240 V AC) ____________________________________ 98
Statement of Conformity (110 - 120 V AC) ____________________________________ 99
Block Diagram – Software ________________________________________________ 100
Instruction Manual – Supplement ____________________________________________________ 101
Autoseeding Unit for Welded High Security Straws (CBS®) ____________________ 101
Introduction ___________________________________________________________________ 102
New Seeding Techniques _____________________________________________________ 102
®
Loading the CBS Straws ______________________________________________________ 103
Position of Autoseeding Unit in Freezing Chamber __________________________________ 105
Part Numbers to Order ________________________________________________________ 106

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Warranty
SY-LAB warrants for one year from the date of shipment each instrument manufactured to be free from
defects in material and workmanship impairing its usefulness under normal use and service conditions
except noted herein. Our liability under this warranty is limited to repair, servicing and adjustment, free of
charge at our plant, of any instrument or defective parts, when returned prepaid to us, and which our
examination discloses to have been defective. The purchaser is responsible for all transportation charges
involving the shipment of materials for warranty repairs. Failure of any instrument or product due to
operator error, improper installation, unauthorized repair or alteration, failure of utilities, or environmental
contamination will not constitute a warranty claim. The materials of construction used in SY-LAB
instruments and other products were chosen after extensive testing and experience for their reliability and
durability. However, these materials cannot be totally guaranteed against wear and/or decomposition by
chemical action (corrosion) as a result of normal use.
Repair parts are warranted to be free from defects in material and workmanship for 90 days from the date
of shipment.
No instrument shall be returned to us without our prior notice (return authorization no. required).
SY-LAB shall be released from all obligations under its warranty in the event persons other than its own
authorized service personnel make repairs or modifications unless such work is authorized in writing by
SY-LAB.
The obligations of this warranty will be limited under the following conditions:
Certain products sold by SY-LAB are the products of reputable manufacturers, sold under their respective
brand names or trade names. We, therefore, make no express or implied warranty as to such products.
We shall use our best efforts to obtain from the manufacturer, in accordance with his customary practice,
the repair or replacement of such of his products that may prove defective in workmanship or materials.
If an instrument or product is found defective during the warranty period, replacement parts may, at the
discretion of SY-LAB, be sent to the purchaser for installation, e.g. printed circuit boards, magnetic
valves, temperature sensors, etc.
Expendable items (e.g. o-rings, seal-rings, indicator lamps, fuses, etc.) are excluded from this warranty
except for manufacturing defects. Such items, which perform satisfactorily during the first 45 days after
the date of shipment, are assumed to be free of manufacturing defects.
Due to the sensitivity of the temperature sensors (Pt-100), the warranty period on these devices will only
be 12 weeks. This applies also to parts made out of PMMA (acrylic glass).
Damage due to any static discharge. Before handling the temperature sensors, we advice the operator to
discharge himself by touching shortly the grounded metal wall of the freezing chamber.
Damage due to lightning, over voltage and/or heavy voltage/current fluctuation on the supply line.

Freezer: 1 year
PC 1 year
LCD Monitor: 1 year
Please Note: For warranty purposes, it is necessary to mention model and serial number of PC and LCD
Monitor on YOUR invoice to the end user.
Do not return any items to SY-LAB without having a RAN (return authorization number).
Note: Shipping- and insurance costs are not covered by SY-LAB
Please read the warranty instructions accompanying the PC and monitor.
In most countries, PC and monitor can be repaired locally.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Purpose
Compact, computer controlled freezing system with integrated, network-compatible PC and connection to
liquid (LIN) pressure vessel up to 22 psia.

This instrument will be used mainly in the field of cryo -preservation of organic materials. With the help of
such instrument, it will be possible to freeze and thaw organic matter under controlled temperature
conditions. This in turn can lead to a higher survival rate (viability) of the material in question after
prolonged periods of storage at cryogenic temperatures. As a cooling media, Liquid Nitrogen at a
temperature of -196°C (-321°F) will be used.

Specification
Freezer
Temperature Range: +40 to -180° Celsius (+104 to -292° Fahrenheit)
Manual and automatic control of heating and cooling
Pt-100 Temperature measurement with 2 (standard) and/or. 3 or 4 optional resistance
thermometers using 4 – wire technique, 1.5 mm Ǿ, 65 mm standard length or 100 mm (optional).
Electrical power-turn-OFF to motor, fan, heater and magnetic valves when lid is opened.
Build-in safety valve – opens at approx. ( 50 psi ) 3,4 bar
Variable LIN conn. from either side of the freezer with original tubing
Monitoring and alarm messaging of supply pressure (optional)
Maximum recommended working pressure: 1.5 bar (recomm. 0,5-0,8 bar)
22 psi (recomm. 7 to 12 psi)
.Autoseeding upgrade possible

Note: Freezer can not be operated above 22 psi - error message shows up on screen when
optional pressure monitoring system is installed.

Cooling rates: 0.01-60°C/min


Heating rates: 0.01-20 °C/min (in range -180°C to -50°C)
0.01-15 °C/min (in range - 50°C to +20°C)
Display accuracy: 0.01°C
Heating power: 750W

Note: The above heating and cooling rates were measured with the freezing chamber empty.

Power Connection
Freezer: (has to be specified on order)
220-240VAC (100-120VAC), 50/60 Hz, 800VA

Minimum PC Config.
Windows® 2000 or XPP, 1x serial, 1x USB, CD-Rom drive and 3 1/2" Floppy disk recommended.
Freezing Chamber
(LxWxH in mm): 217 x 217 x 348
(LxWxH) in inch): 8,5 x 8,5 x 13,7, 16,4 Liter (~4,3 gal.) chamber volume,
Stainless Steel, hinged transparent glass cover, prepared for AutoSeeding, additional sample sensors
possible, thermo fuse over heat protection, optional external alarm outputs.
Dimensions Freezer (LxWxH in mm): 540 x 420 x 520 (w/ PC system)
(LxWxH in inch): 21,3 x 16,5 x 20,5 (w/ PC system
Height with TFT monitor: 740 mm (~29,2 in.)
Height with opened lid: 820 mm (~32,3 in.)
Weight approx.: 29 kg (~63 lbs.)
Required area (LxWxH in mm): 850 x 550 x 870 (depending on configuration)
(LxWxH in inch): 33,5 x 21,6 x 34,3 ( -------------------“----------------)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Dimension Trolley (LxWxH in mm): 710 x 565 x 720


(LxWxH in inch): 28 x 22,3 x 28,4

Capacity Options
1080 (0, 25 / 0, 5 ml) straws or 480 (2ml) vials or 240 (5 ml) vials, 11 metal cassettes or several blood
bags (number varies according on bag size and fill volume)

Software, Data Output


Unlimited number of programs, 12.1“ Touch Monitor (Variant B-D), program menu driven via Touch
Monitor, removal of the heat of crystallization due to counter cooling, Hold and Next Ramp function,
free adjustable start temperature, thermo sensor calibration routine, zoom function, Date- and Time
integrated filename, color inkjet printer (option), program-, chamber- and sample curve.

Normal Conditions of Use


Voltage: (has to be specified upon ordering)
220 - 240 VAC (100-120 VAC), 800 VA
(without monitor, printer and PC)
Frequency 50/60 Hz +/- 2%
Ambient Temp. Range: 15-35°C
Humidity : 20 to 90% relative humidity (at 15-35°C)
Position: Flat and sturdy surface

Transportation and Storage Conditions


If the following mentioned conditions are met, correct operation of the instrument can be expected.

Temperature: +5 to +40°C
Humidity: 10 to 90% relative humidity (non-condensing)
After long term storage, an ice-water calibration might become necessary.

Additional Specification
Protection against memory loss after power failure.
Built in Lithium batteries keeps configuration parameters from being lost during power failure. The
nominal lifetime of such batteries is expected to be around 10 years.

Safety Regulations and Symbols


Terms and definitions used on the instrument and in this handbook.

General Warnings
Warnings signalize the danger of instrument damage, personal injury or even loss of life.

Notes: Notes signalize addition precaution or attention.

Caution, read accompanying instructions

Manufacturer. Mentions name and address of manufacturer.

Date of Manufacture

Serial number of instrument

CE – marking of instrument
AC Alternating Current
Do not dispose with household waste

LIN - Liquid Nitrogen

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Safety Notes
This instrument complies to safety class I – test voltage 1kV (with protective ground).

This instrument was manufactured under following current norms in regard to electromagnetic
compatibility (EMC) for companies in residential, commercial and light industrial areas.

EN 61000-6-1:2001
EN 61000-6-3:2001+A1:2004

Upon request, EMC test reports are available for viewing.

PC’s and peripheral equipment such as monitors, printers, etc. must comply to the norm

EN 60950-1

During operation, maintenance or any other repair work, the following special safety instructions
have to be followed.
If the warnings and mentioned safety precautions in this instruction is not followed, safe operation
cannot be guaranteed.
SY-LAB will not be responsible for any damage or harm violating aboves statement.

Periodic Inspections
Safety-related tests according to ÖVE/ÖNORM E 8751-1 or ÖVE/ÖNORM E 8701-1 or IEC 62353 or
corresponding Norms have to be performed periodically. The test interval must not exceed 3 years.
The tests have to be performed by skilled personnel. For further information on this, please contact
SY-LAB Austria or it’s representative.

Liquid Nitrogen Dewar and Tubing


Liquid Nitrogen supply vessel must comply with the local regulations for pressure vessels. Working
pressure of 0, 3 to max. 1, 5 bar (7 to 22 psi), volume of 60 to 240 liters depending on the number of
freezing cycles. Larger vessels require probably longer supply tubing (available through SY-LAB).
For supply tubes longer than 1.3 m, new cooling and heating parameters might have to be acquired.

Caution: Liquid Nitrogen supply tubes acquired at SY-LAB are made of Stainless
Steel. These tubes remain flexible even at very low temperatures such as Liquid
Nitrogen. If these tubes come with insulation (for protection against burns), the
insulation material will harden at low temperatures and eventually stiffens. In this
condition, the tube / insulation assembly must not be moved at all, otherwise, the
insulation will crack. Just keep in mind that this does not induce any dangerous
situation since the inner part of the insulation, the metal tubing, will stay flexible and
will not crack.

To avoid cracking of the insulation part, let the system warm up to ambient
temperature (takes about 15 to 30 minutes).

Note: To keep the instrument at its highest technical standard, SY-LAB reserves the
right to make modifications / change specifications without the prior notice of the
customer.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Safety

General Precautions
For your personal safety, read thoroughly the following sections before turning the
instrument on for the first time.
Pay attention to all notes, safety and warning messages mentioned in this manual and
shown on the instrument.
Always disconnect the instrument from the power line before attempting to open, clean or
perform any service work. Do not use abrasives or cleaning sprays except when it is
asked for in the manual. Use only a wet soft cloth for cleaning.
Place the instrument on a flat and stable surface (table or desk).
The openings on the freezer housing are mainly used for cooling purposes. Do not block
these openings otherwise, the instrument might get damaged due to overheating. Also,
do not place the instrument close to heat sources or ventilation systems such as
radiators, fans, etc.
Before connecting the instrument to the mains, make sure that the voltage rating shown
on the type-plate agrees with your main supply. If in doubt, contact your SY-LAB GmbH.
representative or SY-LAB GmbH. Austria directly.
Use only the original power supply cable. A grounded (ground potential) wall socket is
required. If the wall socket does not fit the instruments supply cable, contact your local
electrician.
The instrument must be connected to a socket with functioning protective earth
conductor
The electrical environment of the instrument has to comply to the local safety regulations.
In Austria, this is ÖVE-EN 1 or ÖVE/ÖNORM E 8001 (construction of electrical systems
with nominal voltage up to 1000V AC)
When using extension wires or distribution boxes, be sure not to exceed the maximum
power rating of all instruments connected. This is normally about 15A AC.
Never try to insert objects into any instrument openings. Electrical shock might be the
result, which in turn might be harmful to life. Do not pour any liquid into any of the
openings.
With the exception of work asked for in this manual, never try to repair the
instrument by yourself. Only authorized personnel must perform repair work.
For important sample material, we always recommend using an Uninterruptible
Power Supply (UPS).

In all following cases, disconnect the instrument from the mains and call your service
representative:

Power supply cable or connector worn or defective


Liquid was spilled and entered the instrument
Instrument shows malfunction when used according to the operator manual.

Note: Use only accessories (as mentioned in the operator manual) intended to be used
with this type of instrument.

Instrument was dropped and/or housing is damaged.


Instrument shows strange behavior as compared to normal operation.
When instrument was stored for a long time.

Note: For any service requests, always mention the instruments model-, serial- and the
software version number.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Special Precaution (Liquid Nitrogen, Tubing etc.)


Liquid Nitrogen (LIN) is dangerous when it comes in contact with skin or when it gets into your
eyes. Use special care when handling LIN. For safety reasons, you must wear protective gloves,
goggles and a leather apron.
The new model freezers are built with metallic seals and must not be used with any additional
seal rings. If the LIN Connector is damaged, it must be replaced.
During a freezing cycle, the LIN supply tubing is getting very cold and if necessary, must only be
touched with protective gloves. Ignoring this warning might result in severe burns and/or
frostbites.
Areas exposed to excess nitrogen gas (storage tanks, freezers etc.) must be well ventilated.
Under normal circumstances, keeping a window area permanently open (at upper half of the
room) will be inevitable. Nitrogen gas is not poisonous but it will displace the oxygen in the air.
For safety reasons, the use of an oxygen monitor system (available through SY-LAB GmbH.) is
recommended. For badly ventilated areas, it will be mandatory. In any case, please contact your
safety department for local regulations.
National norms have to be considered.
A safety instruction booklet will accompany all supply Dewars delivered by SY-LAB. This booklet
must be read and be understood before handling any liquid nitrogen equipment.
Also, for safety reasons disconnect the instrument from the power line whenever maintenance
work is performed on the instrument. Do not open the instrument while it is connected to the
power-line. Electrical shock might result. We will void all warranty and will free ourselves from any
obligations if our instruments are opened without the prior written consent of the manufacturer.

Precaution for Transportation


During the transportation of the freezer, it is absolutely necessary to secure the Pt-100 temperature
sensors. Use some Styrofoam or similar pad and secure the sensors with tape to the inside wall of the
freezing chamber. This keeps the sensors from bouncing around during transportation.

Caution: For normal operation of the freezer, the Pt-100 transport protection must be
removed. Not doing so will lead to abnormal temperature oscillations.

In addition, do not place any foreign parts or chamber furniture into the freezing compartment.
Damage might occur if done so and will void any warranty.
In case of printer or monitor failure, file a report to SY-LAB GmbH. and contact your local printer /
monitor representative. Most printer / monitor manufactures have worldwide warranty service. Use
only the original packing or pack separately when returned with the freezer. No return shipments will
be accepted by SY-LAB without a Return Authorization Number (RAN). Do not throw the packing
material away. Keep it for possible shipments in the future.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Packing Contents
Please check the contents of the delivered goods. Obvious damage to packing or instrument must
immediately be reported to the shipping agent. Any damage must be recorded on the delivery papers
(invoice, etc.). If the shipment is accepted, then only with "reservation".

In general, if no additional items were ordered, depending on the Variant, it should consist of the
following:

Variant A - Free standing freezing chamber - PC system provided by the


customer

IceCube freezer - basic unit


Power cable
Serial communication cable
Operator Manual
IceCube Freezer Software (CD)
Hardlock (key)

Necessary area for freezing chamber + PC system (L x W in mm): 540 x 400 (without PC)
(L x W in inch): 21,3 x 15,7 (-----“------)

Variant B - Free standing freezing chamber with Touch Monitor

IceCube freezer - basic unit


Power cable
Serial communication cable
Touch Monitor package
Monitor mount
Operator Manual
IceCube Freezer Software (CD)
Hardlock (key)

Necessary area for freezing chamber + PC system (L x W in mm): 740 x 400


(L x W in inch): 29 x 15,7

Variant C - Freezing chamber with Touch Monitor and standard PC-mount

IceCube freezer - basic unit


Power cable
Serial communication cable
Touch Monitor package
Monitor mount
Standard PC mount (two arms) with straps
Operator Manual
IceCube Freezer Software (CD)
Hardlock (key)

Necessary area for freezing chamber + PC system (L x W in mm): 830 x 400


(L x W in inch): 32,7 x 15,7
Note: Max. PC - height: 390 mm (15,4 in.)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Variant D - Freezing chamber with Touch Monitor and mini PC-mount

IceCube freezer - basic unit


Power cable
Serial communication cable
Touch Monitor package
Monitor mount
Mini PC mount (single arm)
Operator Manual
IceCube Freezer Software (CD)
Hardlock (key)

Necessary area for freezing chamber + PC system (L x W in mm): 740 x 400


(L x W in inch): 29 x 15,7
Note: Max. PC - height: 300 mm (11,8 in.)

Optional items for above Variants:

Computer system originally packed


Printer
Accessory package - supply tubing, wrenches, etc.
Furniture f. freezing chamber - racks, seeding, spacer, etc.
Customized orders possible

Note: For monitor, printer and PC package contents, please consult the accompanying operator
manual of the corresponding manufacturer.

Fuses
Two Fuses protect the IceCube against overload. Use only original fuses. The
fuses are located next to the main - power switch of the Freezer Electronic Box.

Value: 2x T 5 A L / 250V
Type: 20 x 5 mm fuse
Fuses
Caution: Disconnect the freezers power supply cable from the mains. Never
change a fuse or perform internal work with the power cable plugged into the
mains supply.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Important Specific Notes


Computer System from third party suppliers must comply with the territorial regulations.
The IceCube in the standard configuration comes with two temperature sensors. One chamber-
and one sample sensor. Optional, two more sensors can be added. However, this cannot be
done in the field. So either you order the IceCube already with additional sensors or the freezers
electronic box is sent to SY-LAB for up-grade.
Liquid Nitrogen Vessels should only be filled to about 90% of their nominal filling volume. The free
space above the liquid level is needed for gas building assuring continues supply of Nitrogen to
the IceCube.
Computer systems not purchased at SY-LAB, the operating system, the freezer software as well
as the Touch Monitor software must be installed. The instructions for loading the freezer program
into your PC are found in this manual. The installation of the Touch Monitor is described in the
monitor handbook.
Since there will be no keyboard connected to your PC, deactivate "Hold on All Errors" in the
computer BIOS. If this is not done, your PC system will stop when looking for the keyboard.
Use only the original Liquid Nitrogen supply tubing from SY-LAB. The original supply tubing
allows the placement of a supply vessel at either side of the IceCube freezer.
For supply vessels up to 70-liter content, the supply tube should be 1 meter long. For larger
vessels, a 1,3 meter supply tube should be sufficient. If other length is required, pls. contact SY-
LAB.
Tubing Connections: 3/4" UNF Nut to G 1/4" conical. Both ends use metallic sealing. No seal
rings are required.
Lock the brakes on the supply vessels wheels in order to avoid unintended movement.
Due to the very low temperature on the LIN supply tubing and connections, water condensation
will occur. This is seen as white fog dropping to the floor. Water droplets in the surrounding air
condense out and will sink to the ground. This is a normal process and has nothing to do with a
leak in the LIN system. Nitrogen gas is invisible and odorless.
Never disconnect a cold supply tube from the supply vessel or the IceCube freezer. Wait
until the supply tube has reached ambient temperature. If moisture due to condensation
gets into the supply tube, it will lead to blockage of ice or to freeze the magnetic valve.
For normal operation of the freezer, the Pt-100s transport protection must be removed. Not doing
so will lead to heavy temperature oscillations.
3 power sockets are required for operation (4 if printer is also used).
The IceCube Freezer must be connected to a computer that complies to EU-norm EN 60950-1
(Information technology – Safety)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 13 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Installation and Check-out

Site Preparation, Space and Clearance


An opening is provided on the backside of the freezer. It is used as an exhaust for the
nitrogen gas. Position the IceCube in such a way that the cold exhaust gas cannot
damage any other equipment nearby. Keep at least a distance of 15 - 20 cm (6-8
inches) to other equipment. A 90° elbow pipe is available optionally to divert the
exhaust (see under “Accessories”).

Additional requirements:

Position the IceCube on a flat, horizontal and stable surface


The instrument must be accessible from all sides
Do not place the instrument such that it is exposed to direct sunlight, extensive heat or cold,
humidity and dust.
Do not use extension wires when connecting the instrument to the power line. These connections
become weak with time and will lead to malfunction of the equipment.
You must be able to remove the power cord from the wall socket at any time. Do not block the
access to the power socket.
Do not use any wall sockets that can be turned ON or OFF by an extra switch or timer. Besides
loosing data when automatically turned off, the sample material might be ruined as well.
Do not use wall sockets where other heavy instruments such as motors, copy machines, ovens,
etc. are connected. Such devices can shortly deplete the supply voltage when turned on.
The computer system and its components should not be placed near big speaker systems,
cordless- and mobile docking stations, etc.
Freezer, computer and all other system components must be connected to the same power
source having the same grounding potential. Potential differences due to bad grounding are
potential sources of error and can lead to the destruction of any connected items. No warranty
will be granted in such cases.

Environment
The instrument will work without any difficulties under normal working conditions (15-35 °C / 60-95 °F).
Nevertheless, in a humid environment, excessive condensation on the LIN-supply-tubing will occur and is
normal. The environment should be such that 90% humidity (non condensing) is not exceeded. Very high
humidity can lead to internal condensation and destruction of the instrument. Also, during transportation,
it is vital that the instrument is given time to adjust to the surrounding temperatures before turned on,
especially in the wintertime, when instruments are transferred from a cold to a warm environment or vice
versa.
Instruments on a trolley must be secured against movement. Use the roller-lock on the wheels to do so.

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Unpacking
Prior to the shipping, the instrument was carefully inspected and packed. Because of the size and weight,
it needs special packing. Unpacking instructions are attached to the outside of the box.
Check for any transport damages before opening the shipping box. If any damage is visible, immediately
contact your shipping / insurance agent or better do not accept the shipment. If in doubt, make at least a
note of the damage on the shipping papers. Take a picture as well.
Check the contents of the package for completeness. Compare with the packing list, delivery note, etc.
If any items are missing, please contact immediately either your representative or SY-LAB Austria. Later
reclamations will not be accepted.
Always state Model Number and Serial Number of
Freezer, Monitor, Printer, and so on.
Do not return any items to SY-LAB without having a
RAN (return authorization number).

Note: Shipping- and insurance costs are not covered


by SY-LAB.

Do not dispose the packing material. You might need it


for future shipments.

On the left, you see an original 14S shipping box.

Size of box (LxWxH in mm): 670 x 520 x 580


(LxWxH in inch): 26,4 x 20,5 x 22,8

Open the box carefully from the top. Below the top, you
will find another cardboard with handles.

Carefully pull the instrument out and pay attention that the instrument does not slip out on one of
the sides.

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Place the freezer on an appropriate location (see "Site Preparation and Clearance").

Depending on the Variant and options of your system, an accessory package will also be included (see
"Packing Contents").

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IceCube Installation - Wiring Variant B:


(Free standing freezing chamber and PC with
and Hook-Up Touch Monitor)
Please consider the above mentioned
environmental conditions. Place the IceCube on This type of configuration includes already the
a flat and horizontal surface. Consider the Touch Monitor.
stability of your table, laboratory trolley, etc. The
IceCube weighs around 30 kg (~64 lb.).

Note: Only trained personnel should perform the


installation of the IceCube.

Unpack monitor, printer, PC, etc. and inspect


visually for any damage. The monitor stand was
modified at SY-LAB (Variant B-D) and does not
exactly look like the one shown in the monitor
handbook.

In the following section, we will go step-by-step


through the instruments installation and hook-up
procedure.

Variant A:
(Free standing freezing chamber - PC system
provided by the customer)

In this configuration, only the freezing chamber


is available. For the system to work, a PC During normal operation, no extra keyboard or
system including monitor and keyboard are mouse will be needed. For hard copies, a printer
required. For hard copies, a printer will be will be useful.
useful.

In addition to the connections mentioned in


Variant A, attach the stereo jack of the PC's line-
The chamber comes with a power- and out of the sound card to the line-input of the
communication cable. Communication will be monitor (not on all units available). Also, attach
established between the freezers serial the monitors Touch-Screen USB-cable to one of
connection and a free COM Port on your PC. the USB ports of your PC. As already mentioned
Use the supplied power cable and attach it to a above, after a successful installation of the
wall outlet. Note: Make sure the freezers power operating system and the IceCube program incl.
rating corresponds to your local supply (see Touch Ware, etc., the keyboard and mouse will
Type-Plate). not be needed anymore.

Thereafter, the IceCube Software can be


installed (see " IceCube Series Software
Installation).

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Thereafter, slide the monitor base onto the stud


Touch VGA Netzteil for the monitor support as shown.
Screen Anschlus Anschlus
connectionA sVGA sPower
nschluss connectio supply
n

(View from underneath)

Generally, the freezer series 14S will come in a


basic configuration (without Monitor and PC
support). Depending on the various types
(Variant A to D), additional accessories will be
included and must be assembled during the
installation at site.
In this configuration, a monitor base will be
included.
Turn the monitor base such that the locking
Installation of the monitor base (Variant B) screw becomes visible. To secure the monitor
Remove the left upper side panel (face plate) as base, use a 3 mm hex driver (included in the
shown. accessory package) and turn the locking screw
counter clockwise (outward).

Attach the monitor base plate to the freezer


frame (4 screws) as shown below. Replace the With the monitor base secured, slide the Touch
left side panel (face plate). Monitor (if already assembled) onto it as shown.

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The viewing angle can be set by adjusting the


swivel head. The electrical wiring is described The viewing angle can be adjusted with the
further above. screw on the swivel head.

Monitor Assembly
The next step will be to modify the LCD Touch
monitor to fit onto the monitor holder

Mounting
Monitor
Holder 2 Plate

Swivel

And finally, the monitor assly. can be placed on


the monitor holder.
Monitor
Holder 1

All necessary parts are shown below. Also, all


required tools are part of this package.

Monitor holder 2 is already part of the IceCube


Variant B shipment.

First, mount the swivel on the mounting plate.

The electrical wiring was discussed further


above.

That completes the hardware installation.


Now proceed to the installation procedure of the
software.

Note: If the IceCube freezer was ordered


from SY-LAB together with a PC system,
the software is already installed and
Then, attach the mounting plate to the back of functional.
the LCD monitor using the provided screws.

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Variant C:
In addition to the configuration in Variant B, this
model comes with a PC support (2 pieces).
Instead of the small monitor base plate with only
one stud as in Variant B, this base plate is
somewhat larger and is equipped with three (3)
studs. The upper stud is for the monitor and the
two lower ones for the PC support.

Replace the left side panel (face plate) and


attach the monitor base as well as the PC
supports in the same way as described above
for the monitor base.

Slide the Touch Monitor onto the monitor base


and place the PC onto the support as shown
(See section above about “Monitor Assly”).

The electrical wiring is exactly the same as


described in Variant B

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Place the Touch Monitor and the PC onto the


support (See section above about “Monitor
Variant D: Assly”).
The installation of the base plate remains
exactly the same as described in Variant B.
The electrical wiring is exactly the same as
described in Variant B.

After the successful installation of the hardware,


the software must be installed onto the PC.

Please refer to the section on "Settings on the


PC" and "Freezer Software Installation" further
down in this manual.

If the PC system was ordered from SY-LAB


together with the freezer, the software is already
installed and ready to be used.

Thereafter, replace the left side panel (face


plate).

The accessory package of this version comes


with a single support for Touch Monitor and
Mini-PC. Attach the support as described earlier.

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When used in conjunction with an IceCube, the


Connecting the LIN Hose to maximum working pressure of the supply vessel
Freezer and Supply Vessel must not exceed 1.5 bar (22psi). At higher
pressures, the operation of the IceCube's
The optional accessory package includes the
internal magnetic valve cannot be guaranteed.
flexible stainless steel hose and a set of
wrenches. The conical metallic seal of the
supply hose does not require any additional
seals. Pt-100 Transport Protection
During transportation, the Pt-100 Sensors
are secured in a block of Styrofoam or
LIN Input
similar material. This keeps the sensors
Max. P: 150kPa
from bouncing around. The sensors are very
(1,5 bar)
sensitive and should be handled with care.
Never bend or drop any sensors.
Do not throw away the Styrofoam block. You
might need it for future shipments.

Liquid Nitrogen Conn.


Metallic Seal

Attach the 1/4" conical end of the hose (small


connector) to the freezer and the other end (3/4"
UNF) to the supply vessel.

Caution: Do not over tighten the nuts. Just


tighten the nuts slightly with the
wrenches. Re-tighten the nuts after a few
minutes of a test run (cooling).

Use only original supply tubes from SY-LAB.


Those tubes have a 90° bent on one side. This
gives the user the possibility to place a supply
dewar on either side of the freezer.

At this point, we again point out the


importance of wearing protective
clothing. All personnel working with the
IceCube must be instructed on how to handle
liquefied gases at sub-ambient temperatures.
Read the safety instructions about handling LIN.

Note: After a freezing run, do not immediately


close the liquid valve of the supply vessel. Leave
it open for another five minutes. Any Liquid
Nitrogen still in the supply hose will boil off and
generate high pressure. If the liquid valve on the
supply vessel is open, excess Nitrogen pressure
can expand back into the vessel. Nevertheless,
a safety valve is built into the IceCube. Any
pressure in excess of about 3.5 bar (51 psi) will
be released to atmosphere.

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supply). Next to the fuses, you will find the main


power switch and the power supply connector as
well as the temperature sensor inputs, the serial
Description of the COM connection and optional (potential free)
Instrument Components alarm outputs.
For replacing the main fuses, a small Philips
and Connections type screwdriver will be needed. The specific
type of Fuse can be found in the section
Before the first turn ON of the freezer, re-check “Fuses”.
all connections.
In the following, we will describe each Caution: Before exchanging fuses or
component in more detail. performing any internal work, always
disconnect the instrument from the
These are: power line (mains).
Freezer Electronic / Box
Touch Monitor (not on Version A)
Printer (optional)
PC-System

Freezer Electronic / Box

There are 2 more openings for Pt-100 Sensors


provided. In the standard configuration, the 2
spare holes are fitted with blind plugs. The
IceCube can be ordered with up to four sensors.
A later up-grade from two to more sensors can
only be done at SY-LAB.
On the Freezer Electronic / Box, you will find the
main switch, power connector, fuses, the Pt-100 Via the COM Port, communication is established
temperature sensor inputs and the COM serial with the PC. The communication cable can
port (9 - pin Sub D) be found in the IceCube accessories. The
connections have to be secured with the
provided screws.
Pt-100
Power
Sensors There are three connections on the Touch
Connector
Pt-100 Monitor (view from underneath):
Sensors
Serial
Power Supply
COM
Main VGA signal and
Switch Touch Monitor USB
Fuses

Touch VGA Power


Grounding Screen connectio supply
wire connection n

The main fuses of the freezer can be found on


the freezer electronic (electronic box / power

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system the next time. Please consult your PC


Establish all connections. Power supply to manual for proper Shut-Off.
power distribution box, VGA and USB Touch
Screen to the corresponding inputs on the PC. After the PC has been shut-off, all other
components can be turned off.
For getting printed results, reports, graphs, etc.,
a printer is recommended. It can be connected Please note: The PC system needs some time
to either the parallel (Centronics) port or to one to boot-up. Generally, the boot-up should not
of the USB ports. take longer than about one minute. Depending
on the settings, the IceCube can be started
Now, all connection to PC and power distribution automatically when entered into the AutoStart
box should be established. list or started manually by clicking onto the
Depending on the type of PC you are using, IceCube icon.
there might be a switched power-output on the
PC.
The monitor normally uses this switched power-
Storage
After every freezing cycle, the freezing chamber
Do not use should be warmed up to about ambient
temperature. This can be accomplished by just
starting a program, which has its start
temperature at about 20°C. During the heat-up,
keep the chamber closed. This has the
advantage that very little condensation will occur
inside the chamber.
Once warmed up, dry the chamber and the lid
with soft cloth or a sponge. If necessary, cleanse
the system with a proper disinfectant.

Caution: Before cleaning the freezing


compartment, disconnect the IceCube
out connector. We would like to point out, that from the power line (mains).
for the operation with the IceCube, this
connector must not be used. Store the IceCube in a dry and dust free
environment. Do not store below +5°and above
+ 40° Celsius. Do not exceed 90% humidity
Turn ON/OFF and Check of the (non-condensing).
System
The freezer system consists of various modules
or components. It is therefore important to follow
a certain turn-on and off procedure. Turn on in
this order:

Freezer electronic
Monitor
Printer (if available)
PC

Important: For synchronization purposes,


the monitor must be turned ON
(especially for TFT Monitors) before the
PC is switched on.

Upon turn-off, it is important to shutdown the


computer system in an ordinary fashion;
otherwise, possible data loss will be the result.
Also, you might have trouble booting up your

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you (e.g. vial- or straw racks, racks for homo-


grafts, bone marrow or blood bags, etc.). If your
system is equipped with an auto-seeding option,
Handling of Temperature various racks for vials and straws are available
Sensors too.
There are typically two temperature sensors in
If only small quantities of samples are frozen,
the freezing chamber. The sensor feed troughs
the freezing chamber is nearly empty; a spacer
are color labeled.
can be used for proper positioning of your
samples closer to the lid. Since the samples are
located closer to the top of the chamber, it will
be easier to visually check the samples (due to
the see-through chamber lid) during the freezing
process, especially during auto-seeding.

As already mentioned, the blue sample sensor


can be inserted into a sample (straw, vial, bag,
etc.). The red chamber sensor can either remain
in the wall socket or be positioned in a well-
ventilated position near the sample material.

The red sensor is the chamber sensor, the blue


one is the sample sensor. When standing in
front of the IceCube, the sensor closest to the
operator is the sample sensor. At all times, the
sensors must be kept free of moisture. The
sample sensor (blue) is used for monitoring the
temperature inside a sample. IceCubes with
additional sensors are available. It can be of an
advantage to measure the temperature of
samples at different points, e.g. in large bags.
The temperature sensors are very thin, allowing
us to insert them into 0.25 ml straws. In addition,
the low mass of the sensor will give us a fast
reaction time.
When used in a vial, never try to force a sensor
into a vial cap. It is necessary to pre-drill or
punch (with awl, nail or small screwdriver) a little
opening into cap. Prepare the opening such that Note: The chamber sensor (red) should not be
the sensor can be pushed easily through the positioned in between sample materials. The
hole. heat released during the crystallization process
Also, never try to pull out a sensor from a frozen of the sample influences the temperature
sample. Damage will be the result. reading and might alter the temperature in the
To remove a sensor from the wall-holder, pull chamber.
the sensor cable slightly out of the feed through.
The sensor can now easily be removed. Once, a program is optimized and working well,
it will not be necessary, to continue inserting the
Caution: The chamber sensor (red) must never sample sensor into a sample, especially if the
be inserted into a sample. This sensor controls sample quantity is limited.
the temperature in the chamber. If this sensor is However, if enough sample material is available
obstructed, the chamber temperature cannot (for record keeping purposes), it will be
properly be controlled. advisable to monitor the freezing process for
every freezing run performed
Various different rack-systems are available Naturally, the sample used for monitoring must
from SY-LAB. Choose the one most suitable for not be used on a patient. It must be discarded.

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In many instances, if the concentration of a


sample in a cryo protectant solution is small, the
cryo protectant by itself (a „dummy“) can be
used for record keeping. Just remember, the
sample sensor will show the temperature within
the sample during the freezing process. In other
words, you know what is going on inside the
sample.

For larger freezing bags, sensors with a length


of up to 200 mm are available (standard length
is 65 mm)

Once more please note:

For normal operation of the freezer, remove


the transport protection from the Pt-100
sensors.

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Freezing Chamber
Description
The IceCube Series Freezer is equipped with a
transparent glass cover (lid). In this way, it is
possible to visually monitor the sample material
during the program run. This is especially
important when determining the point of
crystallization during an Autoseeding run.

At very low chamber temperatures, some liquid


nitrogen droplets are allowed to enter the
freezing chamber but will evaporate quickly off.

Another feature is the hinge. The glass cover


can be opened without looking for extra desk
space. Due to the weight of the lid, the chamber
is firmly sealed towards the top of the freezing
chamber. An opening is provided at the
chambers backside for expelling the gaseous
Nitrogen. As seen from the front, there are two
temperature sensors (standard configuration)
positioned on the left side of the chamber wall.

Straight ahead towards the right side, a plug is


provided for the connection of the AutoSeeding
rack. It is available as an option and can be
installed at SY-LAB upon ordering or be installed
in the field later on.
AutoSeeding Option Kit # (14S): 13-159532

On the right side wall are the in- and outlets for
the air-stream. Nitrogen gas mixed with air will
be withdrawn at the top of the chamber, sweeps
over the heater element (750W) to mix with the
incoming Nitrogen to be expelled at the bottom
of the chamber. So the flow of the air stream is
from the top to the bottom.

The turbulence is generated by a fast moving


fan, dispersing any incoming Liquid Nitrogen to
gaseous Nitrogen.

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Vial rack. It is available for 1.8, 2 ml or for 5 ml


vials. Up to 5 racks can be placed on top of each
other for 2 ml vials and up to 3 racks for 5 ml
Available Freezing Racks vials each holding up to 96 vials.
Vertical Autoseeding rack for either 2 or 5 ml
size vials.

Thus, a maximum of 480, 2 ml vials or 240, 5 ml


vials can be placed in an IceCube 14S chamber.
A maximum of 28 vials can be placed on one
rack. One side of the tubing connection is Note: The freezing racks shown in this section
inserted into the AutoSeeding outlet at the undergo constant refinement and might not look
backside of the freezing compartment. The other the same in the future.
end of the tubing is guided through the large
opening of the chamber and leads to the outside A typical temperature sensor positioning in an
of the chamber. vial is shown. The tip of the sensor should be
inserted to about half of the liquid level.
Caution: Make sure there are no delicate
Instruments behind the Freezer. The extremely It is important that the tip of the sensor is not
cold gas expelled at the rear exhaust might touching the sidewalls of the containment
damage them. (Vial). Keep the sensor centered. This can be
accomplished with a centering disk made of
AutoSeeding rack for straws. It is available for plastic or similar material.
0.25, 0.5, 1.0 and 5.0 ml straws. Additional straw
holders are available.

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A rack for the vertical freezing of blood bags The next picture shows such a non standard
is shown below. This type of rack is adjustable rack
and can be used for a variety of bags.
Size of profile: 184 x 100 mm (7,3 x 4 in.) Rack for 20 ml glass bottles.

It comes standard with 7 profiles and allows the AutoSeeding rack for bags.
placement of 6 bags horizontally (depending on
the size of the bag). If smaller bags are used, a
higher number of bags is possible.

Vertical rack for metal cassettes. Up to 11


cassettes can be place on such a rack.

Manual Seeding for Straws

For special applications, we have the possibility


to produce customized racks as well.

(Symbol photo)

The manual seeding system consists of an


intermediate lid comprising 31 openings for the

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straw holder. Each straw holder can accept up


to 4 straws.

Also, each straw holder can individually be


pulled out of the freezing chamber for manual
seeding. Thus, the temperature in the freezing
chamber remains fairly stable.
The straws can be expelled from the straw
holder easily by just turning the black knob at
the upper end of the holder.

Need a special rack for your samples? Please


ask for a quote. Better, send us a sample of your
containment (bag, vial, glass, beaker, straw,
etc.).

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In the following example, we will show a normally becomes difficult. In order to position
standard vial rack resting on a spacer. This the straws quickly onto the straw rack, we have
simulates a real time situation. The sample made a device, called "Straws Distribution
sensor is positioned in one of the containments, Block".
here in a vial. The chamber sensor remains in
the sensor holder. Depending on the application,
the chamber sensor can be positioned at any
place within the chamber. In many cases, the
chamber sensor is placed next to the sample
material to react as close to the sample
temperature conditions as possible. It is
important that the chamber sensor is not
touching any sample material, vials, freezing
racks, etc. It should measure the temperature of
the nearby airflow.

The Distribution Block is placed below the straw


rack. Then, the straws are placed onto the rack.
Because of the slits in the Distribution Block, the
straws will be equally spaced. Use a small ruler
to straighten out the horizontal position of the
straws as shown below.
Various sizes are shown. The vial rack can be
used for 2ml, 4 or 5 ml vials as well as for many
other vials.

Once, the straws are positioned, the straw rack


can be lifted off and be stacked up to a
maximum of 30 racks / chamber on the 14S,
Please consult our accessories list at the end of each holding up to 36 straws.
this manual for more info on chamber furniture.

One of our furniture is the straws rack. Because


of the amount of straws that can be positioned
into the IceCube14S chamber (up to 1620,
0.25ml or 1080, 0.5ml), the handling of straws

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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

sealed at one end with either powder material,


tampons or with sealing pins (balls, etc.).
Nowadays, many applications require straws to
be welded on both sides to prevent
®
contamination (CBS – High Security Straws).
Instructions are provided in the Appendix.

Here, we show you some of the available


freezing racks, ramps, autoseeding accessories,
etc.

The illustration below shows 3 empty racks


stacked together.

New in our program is the Autoseeding Rack for


Welded High Security (CBS) Straws

New seeding techniques have made it


necessary to re-design the current Autoseeding
system. Previous systems used straws that were

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Program Log-In
Note 1: The surface of the Touch Monitors screen must only be touched with the soft tips of your fingers
or with a soft stylus made out of plastic (such as the ones used on handheld PC's). Sharp and rough
edges can lead to damage of the screen surface. Scratched or damaged surfaces are not covered by SY-
LAB's instrument warranty. Before operating the Touch Monitor, it is mandatory to read the manufacturers
instructions and warranty terms.
---------------------------------------------
Note 2: Initially, the IceCube Series program does not know
which of the Freezers (15M/L-14S) is connected to the PC.
Only after the initial start of the IceCube Series software, the
subsystem of the particular freezer reports back to the
Software and will "mention" its name.
--------------------------------------------
When the IceCube program is first started, you will be asked to enter a User Name and a Password. For
first-time user, the User name and Password will be in both cases

sylab

Use the provided On-Line-Keyboard to enter YOUR User Name and Password if already assigned.

If you do not have this information, ask your IceCube Administrator for it. The Administrator has the rights
to sign you up as a new user. When using the above first-time User Name and Password, the
administrator is advised to change the default User Name and Password from sylab to a real User Name
and Password (this procedure will be described further down).

Here, we enter, (first time users will enter sylab in both fields –
consult the section on “User Administration” to set up a new – real
user)

E.g. Garner Jeff and Password

Press OK to get into the IceCube program or QUIT to revert back


to the desktop.
If you answered OK, a new screen will be presented to you like the
one shown below.

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Description of Program Tools and Buttons


The IceCube program includes several graphical tools. In addition, Windows® based command
structures are available as well. In the following section, we will describe the terminology of the various
buttons, display fields, and so on.

Status Area

Display Area

Main Menu

Instrument
Panel

Logo Area

Operation
Panel

Main Menu Pop-up menu. Behind this menu button, sub menus are found for File, Display Options,
Settings, Version information and Maintenance.

Please Note: The access to various pop-up menus depends on the USER-LEVEL granted to you by
the IceCube Administrator.
Here, we assume that all levels (administrator level or when initially logged in with sylab - sylab) are
granted.

In the File menu, patient information, freezing programs, freezing runs and reports can be generated or
edited as well as the data and program path (directory) be set.
In Display Options, one can choose the way the program curve will be displayed. In Total View, the
whole temperature range +40°C to -180°C can be displayed. Tracking view follows the program
temperature and shows a somewhat narrower temperature range.
In Settings, the parameters for X-axis (time) and Y-axis (Temp.) can be defined, remaining or elapsed
time be shown on the screen, the current program temperature be displayed next to the temperature
curve and the pressure curve be displayed. A more detailed description can be found further down in this
manual.
In Options, some sub menus pop up. In Device settings, various alarms can be turned on/off and the
pressure alarm limits be set. Users can be administered, giving each user his own Name and Password.
Finally, the language can be selected. At the moment, only German and English language files are
available. More language files will be available in the future.
Version Info gives us some information on the installed PC-program version as well as the version
number of the instruments resident program. In case of failure, this information together with the
instrument model and serial number should be given to the service personnel.
Maintenance allows the user to perform an Icewater calibration as well as to set the correct
Communication port

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Status Area Display of the current instrument action like heating or cooling ON or OFF, alarm 1 / 2
status, ventilation system running, online- (CTS) and text status, etc. All statuses are displayed in form of
signal lights. Is the light ON, is the corresponding function active.
Display Area Graphical display of program-, chamber-, sample temperatures, and so on. During
the program run, the temperature curves are displayed.
Instrument Panel Numerical display of program-, chamber- and sample temperature(s) and remaining
time or run time of a program. The display for the 3rd and 4th sensor is only active if these options are
installed.
Operation Panel Buttons for starting and stopping a program run, initialization and triggering of
manual seeding, Hold and Continue button, Program Load button.
to StartTemp brings the freezing chamber to the pre programmed start temperature. Seeding (option)
opens an additional magnetic valve, Hold keeps the freezer suspended at the current status, Next
advances the program run to the beginning of the next step and Stop stops the freezing cycle.
Logo Area Above the non-removable SY-LAB logo, it is possible to add an additional logo, i.e.
your own, if desired. A bitmap (.bmp) format with a size of 106 x 122 pixels should be used.

In the following, we will describe in detail the various tools and buttons.

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Menu Area
Main Menu - File

Under the Main Menu - File, six sub menus are


shown as seen on the left.
The Edit description will allow us to enter sample or
patient specific information.
Open historical data allows us to locate, view and to
some extent modify already finished freezing process
data. On the Validation Software package, all
modifications are recorded and are traceable
In the Freezing program, new programs can be
written or existing programs be edited.
The next two entries regarding data and program
directories give you the possibility to select your own
location (directories) where data and programs might
(will) be saved.

Exit will close the IceCube program.

Edit description
As already mentioned, on this screen, information about a sample or patient can be entered.
The first field will accommodate the Freezing Program Name. You will be able to reach this screen either
by selecting Edit description just described above or it will pop-up just before the actual program run is
started.

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If the Edit description was selected before loading a program, no Freezing program name will be
displayed. The program name will be automatically entered as soon as you load a program later on.
However, information about a sample, patient, operator, etc. can be entered in the following field lines.
The Last field will ask you for a sample or patient name.
This field will be divided into a Last and First name area. It is important to enter some specific
identification in the Last field. This will help you later on in finding a specific sample or patient in your data
file directory.
So Last will require some input. We would suggest entering the last (family) name of a patient or the
name of a specific sample. The first 8 characters of Last will be part of an automatically generated
Process record file name. The remaining characters of the name will be truncated. All other field entries
except the operator name are not required but are provided to enter other useful information such as
Sample or Patient ID and Memos. The Operator name will automatically be filled in and will be identical to
the Login name of the currently logged in user. If more than one user is operating this instrument, they
must each have a separate User Name. Pls. consult your system administrator to set you up as a new
user.
On the bottom of the Edit description screen, the automatically generated process file name can be seen.
As you can see, the first part consists of the Last name. In addition to the Last name it will include the
year, month, day, hour, minute and seconds. In this way, there can only be one specific file name for
each freezing process. As an example, the file name might look like this:
Entering any information into the various fields is accomplished with the pop-up keyboard.

Garner_2003-03-18_13-47-26.prc

This corresponds to the file name convention

LAST_yyyy-mm-dd_hh-mm-ss.prc

The extension of the file name PRC indicates that it is a freezing PRoCess file.

As already mentioned earlier, this screen will also be presented to


you when you press the "to Start Temp" button after loading a
program. Keep in mind that in order to save a freezing process
record, we have to give it a specific name. Therefore, you cannot
start an actual freezing run before giving it a Last name. You MUST
enter something in the Last field.
Also, use for the Last name only characters which are allowed
under the Windows® file name convention. These are all characters A-Z, a-z, 0-9. Do not use special
characters such as / \ < > , . ü,ä, etc.

Two Memo fields are available for adding additional information. These two fields can be edited even
after a process file run whereas all other fields are protected against any changes. Just keep in mind,
after a freezing run, the operator, patient name, ID, etc. must not be changed for safety reasons.
However, some additional information, e.g. about the freezing run might need to be recorded (in the
Memo field).

Note:
In the validation version, any changes in the Memo fields are traceable and can not be deleted.

On the right of each memo field, a message will let the operator know of how
many changes were made to this particular memo field. If modifications were
made, "Show" will list all changes that were made in the past. It will show who
the operator was and at which time and date the "editing" occurred.

In our example, it will look like below. Zero "0" modification(s) has changed to
one (1) showing that one modification was done to the particular memo field.

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Please keep in mind that the “Show x modifications” buttons are not present in the
standard IceCube Software.

As soon as the requested information is entered, you must press OK

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Open historical data


For viewing freezing process records, we have to get into the Open historical data menu. Either a clear
freezing process screen is displayed or a previously loaded process record is shown.

On the right bottom of the screen, the Menu button gets you into various sub menus.
At this point, Show description will display the patient or sample information like in Edit description above.
Show program will display the original freezing program if one was loaded with Open process record and
Display Total View will show both, the
program curve as well as the associated
freezing run.

In the File menu, you can choose among

Process directory, Open last run, Set Source


directory, Print Diagram, Print Program &
Events, Print both of above and Close
window.

Process directory
When Process directory is selected,
you first have to select a process record
from a file list, assuming that you have
made a freezing run previously. If not,
this list will be empty and no historical
data exists.

Open last run


Open last run will display the last freezing run that was made.

Set source directory


Set source directory allows you to choose the location where you want to store the Process record files.

Print Diagram, Print Program & Events, Print both


Print Diagram, Print Program & Events and Print both of above will produce a printout of the process
curve, process curve and a tabular listing of events ( Hold, Continue, Next, etc.) or both of the previous.
Before printing starts, a preview will be shown on the screen.

Close window
Close window will allow you to exit this page and to revert back to the previous screen.

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Let us go back to the Process record for a more detailed discussion.

If this menu was chosen and a process file opened (e.g. Testcurv_2004-07-....), following screen will be
displayed.

The process record will then be displayed, can be enlarged, etc. and be printed out.

The above curve is a typical freezing process test curve to


find the temperature of crystallization. However, here we can
see clearly the program temperature curve (green), sample
temperature curve (blue) and hidden behind the green curve
the chamber temperature curve (red).
The area of interest can now be zoomed in (to enlarge the
display area, draw a rectangle around it from left upper to
right lower corner) and be Added to the List on the right. You
can choose as many windows as you like. When added to
the list, the window can be printed out later on.

In the right area field, you will see the windows information.
Hier, 23.96 -4.13 44.52 -30.00

It means that the left upper corner of


the selected window is at 23.96
minutes and -4.13 °C. Whereas the
right lower corner of the window is at
44.52 minutes and -30.00°C.
Choosing "Events" on the above screen, additional Information about each
"Action" performed during the program run can be displayed.

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If this is all we want to print out, press Menu - File - and choose among the various print options.

Print Diagram: As the name implies, this will only print the
diagram(s)
Print Program and Events: Will print all the program steps and
events like opening and closing the lid, pressing manual seeding,
stop, etc.
Print both of the above: Again, as the name already implies, both
of the above, diagram(s), program steps and events will be
displayed on screen and eventually be printed if you desire so.

If you choose Print both of the above, the result might look something like
the display below.

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List of Available Tools


The size of the graphical
display can now be
modified using the three
buttons shown.

The arrow buttons work like buttons on a tape recorder. The two middle
buttons move through the various pages, page by page, whereas the two
outer buttons move the display either to the first page or the last page. Some
of you might have only one page to display. In this case, the arrow buttons
are not used.

The printer sign on the left is used for the printer - setup. The diagrams can be
printed out locally or be sent to a remote (network) printer.

A message (description) appears when the cursor is positioned over an icon.

Close reverts back to the main screen.

Show description, Show program, Display Total View


In the sub-menu Main Menu - File - Open historical data - Menu, we have the possibility
to look into the Freezing Prozess Information (Show description), look at the original
program curve (Show program) and display the complete program curve and process
file (Display Total View). Naturally, a process file must be opened first, using File -
Prozess directory (choose your process record) or Open last run in order to display the
last freezing process data.
We have now briefly discussed the Edit description and Open historical data menus.
The next menu Freezing program will allow us to write a new or edit an existing program.

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Create / Edit freezing program


As the name already implies, here we can create a new program or edit an existing program. Actually, we
are getting again in the program edit screen as seen below.

When Create new is selected, a new program with only


two steps (0 and 1) will initially be displayed. Step #0
represents the start temperature. It is the temperature, at
which the sample should be loaded into the freezing
compartment. In many applications, this will be either 20°C
for ambient temperature or about +4°C for refrigerator conditions. The
default starting temperature value is +20°C.

Also, as a default, the slope in step #1 (temperature gradient) will initially be set at -1.00°C/min.
In order to edit an existing program, we would choose File / Edit existing and select the program we
would like to edit.

More on this, further down in this manual.

Set data directory


The process file of a finished run has to be saved somewhere. Here,
we have the possibility to choose an appropriate location. For clarity
and ease of locating your data later on, we suggest putting the data
sub directory in the same location as the IceCube freezing program.
Assuming that you installed the IceCube program in a directory called
Freezer, a data directory called Process Records will be created.
In this Process Record directory, all future process records (data) will
be saved.
Note: In order to prevent data from being lost, do not forget to backup
your process records periodically.

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Any other directory can be used. Also a new directory can be created.

Note: Some expressions appear in German language. This is due the German Windows®
Operating System installed. These expressions are replaced by similar words depending on
the Windows® language version used.

Set program directory


As already mentioned above in Set data directory, the same applies here. You
simply can choose the location where to store your freezing programs. Again,
we suggest storing the freezing programs in a sub directory of your IceCube
program directory.
But again, any other directory can be used. Also a new directory can
be created.

Note: To avoid loss of data and programs, it is advisable to make


occasional back-ups.

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Exit
The last file menu entry is Exit. As in most other Windows® based
programs, Exit will close all processes within the program and reverts
back to the desktop area. All running processes must be stopped or
finished otherwise, you will not be allowed to Exit the IceCube program.

Note: To avoid loss of data and programs, it is advisable to make occasional back-ups.

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Main Menu - Display


In this menu, we can set the way the program curve will be presented on
the screen during a freezing run. Three sub-menus are available, Total
View, Tracking View and Settings.

Total View
Total View will give us an overall view of the program temperature. This is
currently between + 60°C to -200°C maximum. However, if the last step in
the freezing program ends at e.g. -120°C, Total View will show a screen
with the lower temperature limit set at -120°C.

Tracking View
Tracking View will follow the temperature curve as the program run progresses. This will give us an
optimal presentation of data on the display.

Settings
The remaining Menu entry is Settings.

Setting allows us to define how the screen behaves during a freezing run. A freezing run may need a long
time, thus requiring drawing curves beyond just one display screen. So we have to
tell the program at what stage to update the screen contents for both temperature
and time axis.
In addition, we can choose if we want to get the elapsed or remaining time
displayed in the instrument panel.
Also, we can choose to get the temperature displayed on one of the temperature
curves.

Time axis
On the left side of the screen, the time axis can be set. In our example,
during a program run, a time window of 18 minutes will be shown. When
the temperature curve approaches the right edge of the display area, just
0.5 minutes before, the screen will shift to the right by 1 minute.

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Temperature axis
The setting of the temperature axis functions somewhat similar. However, the up-shift as well as the
downshift can be set independently. In our example, the display range is set to 40°C. When the display
reaches the upper or lower edge of the display area, the up- and down shift adjusts the screen by 10°C.
In this example, the up- and downshifts are set equally. Under normal operating conditions, it is not
necessary to change these settings.

Time elapsed / remaining

The second instrument from the right on the instrument


panel is showing either the remaining or elapsed time of
a program run depending on the setting above. Just
mark the button show elapsed time or show remaining
time as desired.

Actual program position


During a program run, the current temperature of the program temperature
curve can be displayed just above or below the curve. When Actual
program position - Display numerical temp is marked, a little circle
appears right at the tip of the program temperature curve.

Temperature value to track


Since we marked - Temperature value to track -, the screen will focus on
and will follow the e.g. Chamber temperature

LNi pressure
LNi pressure - show curve is not implemented in the 14S.
(Can be added as an option – factory installation required)

If all settings are done, press OK to revert back to the previous screen.

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Main Menu - Options


In the Options submenu, the Device Settings can be changed and
saved, the Users be administered and the Language be selected.

Device Settings and alarm configuration:


To get into this menu, select Main Menu - Options - Device Settings.
Following screen will be displayed.

In all cases, you can see that


besides an internal alarm,
additional alarm outputs can
be set. However, this requires
additional hardware within the
freezer. The additional alarm
outputs can be ordered as an
option with the freezer directly
or be upgraded later on
(freezer box must be sent to
SY-LAB). The alarm-relay
outputs provide potential free
contacts rated at 250VAC /
10Amp.

Here, we can set the condition


and duration of various alarm
outputs on the IceCube Freezer.
In short, these are:

Indication that start temperature has been reached


Alarm outputs when:
temperature deviation is out of the set limits
defect of temperature sensor
a critical power failure occurred
a short time power failure occurred
the freezing program is finished

Note: LNi pressure Alarm not activated on 14S units

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Description of the various conditions and alarms


Note: All displayed values are the default settings

Start Temperature reached condition


On the right, you can see the corresponding display-field.
Here we can determine how a start temperature reached
condition is defined. In our example, if the start temperature
of the freezing chamber (defined in the freezing program) is
within +/- 0.5°C and if this condition prolongs for at least 20
seconds, the start temperature reached indicator will light up.
As you know from the above description, the "to Start Temp"
button will change to "Start" when the program is started. But when the program reaches the start
temperature and meets the conditions shown above, the "Start" buttons will be illuminated. This functions
only once. As an example, if the start temperature is reached, the button lights up. If however, START is
pressed, the light is turned off. If the cover is now opened, the temperature now rises above the start
temperature and returns back after the cover is closed, the Start button will not be illuminated again.

LNi pressure Alarm (not implemented on 14S)


During a freezing run, the pressure in the supply vessel will be
monitored, thus giving us some information on how a freezing
program behaves when the pressure changes. Here in our
example, an internal alarm (Buzzer) will be triggered when the
pressure falls below 0,1 bar or when the pressure exceeds 1,5
bars. Besides getting a corresponding message on the screen
(when the PC is operational), an acoustic alarm will be heared.
The appearance of this acoustic alarm depends on the settings
of the four time fields. In our example, we turn the buzzer ON
for 1 second, and then we turn it OFF for 1 second. Again we

turn the buzzer ON for 1 second but now,


we turn it OFF for two seconds. So this
alarm will have a distinct sound, like

Beep (1sec.) - Break (1 sec.) - Beep (1


sec.) - Break (2 sec.)

Note: In the following alarm descriptions, the internal buzzer alarms will behave in the same way as just
described and will not be mentioned anymore.

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Allowed Temperature deviation


Whenever the actual chamber temperature deviates from the
program temperature by more than the pre-set value, an
acoustic alarm will be issued according to the settings on the
right.
Here, the internal buzzer will give an alarm whenever the
program- and chamber temperature deviate more than +/- 5°C. If
the temperature deviation is getting again within the specified
limits, the alarm will be resetted.

Temperature reading invalid


The readings of the temperature sensors are always compared
with values in a tabular listing.
If the readings are wrong or out of some limits due to a sensor
short or break, a visual as well as an acoustic alarm will be
issued according to the settings on the right.

Critical power fail - awaiting user assistance


As already mentioned earlier, a critical temperature window can
be defined. This will be most likely the temperature range in
which crystallization of the sample takes place. In case, a power
failure occurs within this critical temperature window, the alarm
conditions on the right apply. In addition, you will get a
corresponding warning message on the screen, when power is
re-established to the PC.

The warning message also will list the time at which the power failure occurred. It is now important to
determine in which state the sample is. According to the sample condition (frozen or not), the operator
has to take an appropriate action. Either to discard the sample, re-run the freezing procedure or placing
the sample into Liquid Nitrogen

Also, another window will be displayed. The Re-Start Point


window. Here, the user has now the possibility to determine where
the program should continue with the program run. Either time or
temperature can be selected. It is also possible to set the re-start
point graphically by positioning the cursor at the point of restart.

Short time power fail


A short time power fail is a non critical alarm. Since the power
failure occurred only for a few seconds, we can assume that
this did not harm the sample. As soon as power is re-
established to the freezer, the freezing program will proceed
as if nothing would have happened. Some hospital power

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supplies will take a few seconds to switch over to the auxiliary supply. So the IceCube Series Freezer can
handle such systems and continues to work. Nevertheless, an alarm condition is issued, acoustically as
well as visually. The visual warning message will be seen only when power is re-established to the PC.

Freezing program finished


At the end of a program run, most operators would like to get
notified (e.g. to transfer the sample(s) to Liquid Nitrogen). For
this purpose, we are using the program finished alarm. It will
give an alarm but can be set such as to give alarm on the
internal freezer buzzer as well as on the local PC.

Save Settings
To keep those changes also for the future, it is necessary to Save the Settings in
memory.

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User Administration
One of the menus in the Options menu is "User administration". User administration allows the creation
of a list of users. Each user has to have a specific name and password in order to Log-In into the IceCube
Series program. Depending on the user rights, one has access to functions that others do not.
Initially, IceCube Series software is delivered with the administrator log-in name and password set to
sylab.
We strongly recommend to change these settings top your personal User Name and Password.

Note: Before deleting sylab, make sure you have tested your personal log-in.

The following screen opens if logged in as an administrator with all rights

or this screen,

when logged in as normal user, having no administrator rights.

Note: Again, depending on the settings of the user rights, the user will have only access to functions for
which the administrator gave permission to use them. Initially, when Logging-In as sylab , administrator
rights are granted. The administrator should then create his own User Name and Password, create for
himself administrator rights and set up other users with limited or equal rights. The user sylab should
then be deleted.
First, you will be presented with a list of users if they were previously entered. To create a new user,
select New user. The keyboard pops-up and you will be able to enter a user name, password and
password verification. Each field needs at least 5 characters.

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Just press OK to accept the new "USER". If you have not entered exactly the same characters in the
password verification field as you have entered in the password field, an error message will be displayed

"password and verification do not match"

Correct the problem by typing in the same information in both fields. Also, if the user name or the
password is too short, it will let you know "minimum required length is 5 characters".

In addition to enter a new user, users can be deleted or passwords be changed.

Once you have entered a new user, Close form to revert back to the main screen.

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Info for the Administrator

Administrators have the right to grant or not to grant other users access to various functions.
Following functions can be selected or de-selected.

These are:

Look at previous freezing runs-----------------------------

All available programs can be used ----------------------

Start the act. prog. before the start temp is established

Stop freezing run before the program is at the end ----

Allow the user to use the HOLD/CONT. button-------

User has permission to jump to the next step----------

Give permission to turn on manual seeding--------------

Not available on 14S (Trigger) -----------------------------

Allow the user to write or edit a freezing program -------

Permit the modification of the two memo fields ---------

Change the path of the data directory ----------------------

Change the path of the freezer program directory -------

Choose another language ------------------------------------

When changes are made, allow to save them perm. ----

Allow the user to make changes to the device settings -

Let user perform an ice-water calibration -----------------

Give permission do change load device settings ---------

Allow the user to update the subsystem of the frezzer -

Give permission to use a different communication port

Grant administrator rights -----------------------------------

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Language
In the IceCube Series software, you will be able to select among one or more
different languages. At the moment, only German and English are available.
Other languages can be added if a need for it arises. The language file name
ending will be *.LNG. For example: Engl117b_1.LNG

When you open the language menu,


you will be asked to choose the
directory location and the name of the
language file.
Please note: Once you have selected a
different language file, it is necessary to
close and re-open the IceCube 14S
freezer program.

Using the new language file as the default, it will be necessary to press "Save Settings".
Next time the freezer is turned on, it will have the new language file setting.

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Main Menu - Version Info


Behind this menu, we can find the program version of the currently installed IceCube
program. This information together with the serial number of the instrument
will allow us to search through our production records. It is like the fingerprint
of your instrument.

Main Menu - Maintenance


There are two submenus in Maintenance, Ice water calibration and communication.

Ice-Water Calibration
From time to time, it might become necessary to perform an ice-water
calibration. Electrical components are slightly changing their
characteristics with time. So it is advisable to occasionally re-calibrate
the temperature sensors.

To check if calibration is necessary, immerse the sensors into an ice


water slurry, stir for a few seconds and monitor the temperature
readings in the instrument panel. If the readings are close to 0,00 +/-
0,04 °C, then no calibration is required. If the readings deviate more
than +/- 0,04, re-calibration should be performed.

In any case, if a temperature sensor is


exchanged, an ice water calibration must be
performed.

Get some crushed ice and pour it into a mid-size coffee cup. Add some tap- or distilled water to it. Stir
thoroughly to get a good mixture of water and ice (slurry).

Now take the temperature sensor(s) out of the


chamber holder and place it into the slurry. During the
calibration, slowly agitate the sensor(s), making sure
that the temperature distribution in the cup is constant.
Do not touch the walls of the cup. Also, hold the
sensors as shown in the picture in order not to
influence the temperature readings. Agitate for about a
minute to reach thermal equilibrium.

Enter the Ice Water Calibration screen by selecting


Main Menu - Maintenance - Ice water calibration.

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The calibration screen opens, showing you a lot of


temperature readings and digital data.

You do not have to enter or change any of the


values you see in the tabular table.
The only functions we can use on this screen are:

Calibrate
Load from ....
Save as ....
upload

When you receive the instrument from SY-LAB, you


also will get a diskette with calibration data. These
are the default calibration parameters for YOUR
freezer. They are stored in the freezers memory
already and are provided only for the purpose of
having the default parameters available in case
somebody overwrites the internal calibration
parameters with erratic data (e.g. performing an ice
water calibration at ambient temperature).
If the calibration parameters are lost, they can be
reloaded, using the function Load from ...

When the freezer undergoes a new ice water calibration, the new calibration data should be saved. This
is accomplished by the Save as ... function.

The actual calibration is performed with Calibrate.

Now, let us continuo our discussion from before. With the particular sensor in the ice water slurry, press
the corresponding correction button at the bottom of the screen. The first field is for the Chamber
temperature sensor, the second field for the first Sample temperature sensor (other temperature sensors
are optionally available) and so on. When you feel that the slurry is agitated well, press the Calibrate
button.

.../..Chamber Sensor..../....Sample Sensor...../....Additional sample Sensors.../

The reading from the temperature sensor at 0°C is then compared with the tabular listing. The difference
or the correction value will then be displayed in the corresponding field in °C.
After calibration, the new calibration data must be uploaded to the electronics. Press upload. Also, store
(Save as ... ) the new calibration data either on Disk or write it to a sub -directory for later retrieval if
necessary.

Communication
In the communication menu, we can set, as the name already
implies the communication port (COM-Ports).
Depending on the number of COM-Ports on your computer, you
will be able to select among COM1 / COM2 / / COMn.

Select the correct COM port and "Save Settings".

At this point, we will continue with writing a little test program to get you started.

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Writing a Test Program


Note: The following description assumes that full administrator rights are granted.

Test program for finding the crystallization temperature of a


sample
In this section, we will describe in detail how to write a new program. At first,
we will use a typical test program for finding the crystallization point of a
sample. Keep in mind that for every individual sample material, a new program
curve must be found. In our example, a 2ml vial, filled with 1ml of a 10%
Glycerol / Water solution will be used. Since this will be a fairly easy program,
it is perfect for getting the hands on the equipment. Following the test curve
programming, we will use the results from the test curve to create an actual
freezing program curve.
As already mentioned, we will try to find the crystallization temperature of a
10% Glycerol / Water solution. For this purpose, we slide the tip of the sample
temperature sensor through the cap of the 2ml vial. Getting good temperature
distribution results requires the tip of the sensor to be in the geometrical center
of the solution. The sample temperature sensor (blue) is the one closest to the
front of the freezer.
Now we can start creating our first (test) program. For this purpose, we will
focus on already existing programs. The start temperature of the freezing
process depends in most cases on the initial temperature of the sample or freezing (cryo) protector. Is the
cryo protector at refrigerator temperature - (about 4-5°C) and the sample + cryo protector kept at this
temperature, then, our freezing process should start at this temperature. However, is our sample / cryo
protector at ambient temperature, then we will start the freezing process at ambient temperature - about
+20°C.

In our example above, we will start the freezing process at +20°C. Then, we will bring the cryo solution
(sample + cryo protector) down to zero (0°C). At zero degrees, we will remain there for some time to give
the cryo solution time to equilibrate at this temperature. In other words, we give the cryo solution time to
get close to the programmed temperature. In this way, we always start with the same sample temperature
conditions. So we will cool the cryo solution with -2°C/min. from +20°C down to 0°C. Then, we will keep
the temperature at zero for 5 minutes.

Please note: This is only an example. Cooling rates, equilibration time, crystallization, and so on might be
different for your sample material, cryo solution, etc.

The point of crystallization should be found while slowly cooling the cryo solution. As slower the cooling
takes place, the higher the chance that we find the exact crystallization temperature. However, if the
cooling (slope) is too slow, the program will have to run for a very long time. This in turn will lead to a high
Liquid Nitrogen consumption. So we have to find a compromise. A cooling rate of -1°C/min. is widely
accepted and used in many applications for approaching the crystallization temperature. From
experience, we will assume that the crystallization temperature of our Glycerol / Water solution will be
somewhere around -7°C to -10°C. So in order to cover this temperature range, we will program a lower
temperature of -40°C. In other words, we will have a temperature step going from 0°C with -1°C/min.
down to -40°C. This is all we need for our test curve.

In summary, the program, let's call it TESTPROGRAM will look like this:

Step Temp - °C Slope - °C/min. Time - min.


0 20,00 0,00
1 0,00 -2,00 10,00
2 0,00 0,00 5,00
3 -40,00 -1,00 40,00

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With the information above, we will now program our first freezing process. For this purpose, we have to
get into the Main-menu - File and select Freezing program - Create new.

Freezing program editor

Edit control - Program


The following screen will be opened.

Edit Freezing program points


Initially, one temperature segment (step) - +20°C to +10°C, with a slope of -1°C/min. is
displayed as a default. At first, the program segment will be displayed as a green line.
We have now various possibilities to enter our program. One would be to graphically
move the end points of a temperature segment around.
This is OK for getting an approximate program curve.
Another method would be to use the arrows next to the temperature, slope and time
fields.

The various steps are then shown in a tabular listing at the upper right of the screen.

However, entering an exact program will require the input through the keypad. In this way, all entries for
Temperature, Slope and Time can be entered exactly as we want them. To
open the keypad, just click in the Temperature, Slope or Time field.
Immediately, the keypad display opens. A second double click in one of the
fields will close the keypad again.

The Temperature, Slope and Time values can now be changed numerically by
typing in a value or values can be changed by using the up and down arrow
buttons.
The left arrow is used as a back-step button (removes character by character
to the left of an entry). In addition, there is a delete (del), clear input, enter and
minus (-) button.

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Now we want to keep the first temperature point


at +20°C. The end-temperature point however
should be at 0°C with a slope setting of -2°C/min. Touch the green end
point of the first segment if it is still green. The green line will turn to
red. Now, double click in the temperature field. The keypad opens.
Enter 0,00 and press Enter. The value in the temperature field will
change to 0,00 and the Slope field will be highlighted next. Type in -
2,00 for the Slope and press Enter. The Time will be adjusted
accordingly. Here, it will display 10 minutes. Naturally, a temperature
difference from +20°C to 0°C with a speed of -2.00°C/min. will take 10
minutes.

Let us enter the next step in the program. Press Append Point. The
previous segment (step) will turn green and the new step will be red.
Repeat the procedure above. This time, we would like to remain at 0°C for 5
minutes. So we set the temperature to 0.00°C. Click into the temperature field and
enter 0.00 on the keypad. Then, click on time, press clear input and then type in
5.00 and press enter.

Again, to add the last step, press Append point, enter -40°C for temperature and -1.00°C/min. for slope.

The display should look exactly like the one above. If you have made any mistakes and you want to
change any of the steps above, use the Previous, Next, Delete point or Insert point buttons to correct
them.

Previous will move you back in the program. Once the first step is reached, the Previous
button will be disabled.

Next will advance step by step to the right until the end of the program is reached. On the
last step of the program, Next will be disabled.

Delete point will remove the red (current) program segment from the screen and also from
the program list on the right. Caution: If you have pressed Delete point by mistake, you
have to re-insert this program segment.

Insert point will add a program segment just before the current step (red). If step 3 is the
current step, Insert point will add a step just between the second and third program
segment.

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Append point will add a new program segments to the end of a previous segment. So
Append will add new steps to your program.

Now it is time to save our little test program.

Filing operations
The four buttons on the right are used for filing operations.
File/New will clear the screen and you are ready to enter a new program as described
above.

File/Save will allow you to save your freezing process curve (program). You will be asked
for a file location and file name if it is a new program. If you opened an already existing
program, file save will re-save your file under the same name without asking for another
name.

File/Open will allow you to open an already existing program. It can then be edited and saved under the
same or a different name.

File/ save as will give you the opportunity to save a new or existing program under a different file name.

File/ Exit closes this screen and reverts


back to the Main-Screen

Press File/Save and enter TESTPROGRAM


as file name. If this name already exists, you will be asked to either enter another name or to overwrite
the existing name.

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Axis standard
Axis standard will always display a default screen width of 180 minutes and a screen
height of 170°C (+30°C to - 140°C).

Mouse button
In order to provide the function of a right mouse button, one can press on the
standard mouse symbol. It will change its appearance and can now be used
once as a right mouse. If this function is needed repeatedly, just press on the
mouse symbol.

Active point size


Active point size - will allow you to change the size and sensitivity area of the movable pointer (red
dot). The larger pointer can be moved around more easily on the touch screen and is provided only for
your convenience.
Before we start the freezer to determine the crystallization temperature of our sample, let us discuss the
way windows or screens are zoomed in and out, screens moved around and so on.

Moving Screens and Zoom Functions


Program curves can be very long and might not fit on one display screen. In order to move
from screen to screen, a mouse button was installed on the program screen. When the Touch
Monitor is touched, the function of the left mouse button is activated.
In order to enlarge an area of your screen, you can draw a square around this area by moving
from the left upper point to the right lower point just as you would with a mouse in a
Windows® based environment
To repeat, an enlargement can be accomplished by drawing a rectangle starting at the top left down to
the bottom right.

If we reverse this procedure, starting to draw a rectangle from the lower right corner to the top left corner,
we have reset the screen to the Total View. Try it out. Do not hesitate, move your fingers over the screen
and test a little bit.

Now as already mentioned, this would be the function of the left button on a standard mouse.
However, for some features, we will need also the function of the right mouse button. No
problem. Just press the mouse button on the lower right of the screen. The mouse image will
change (red right button) and will be active for only one mouse action. Thereafter, the image
changes again back to the left mouse button.

When will the right mouse button be used? Whenever you want to move the whole screen around.

If the right mouse button is active, touch the monitor and move with your finger around. You will see that
the screen is shifted to whatever direction you move your finger to. In this way, it is possible to move
around on the screen for optimum display comfort.

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Running the Test Program


In the previous section, we have learned how to write a simple program. Actually, this program can be
used for finding the temperature of crystallization of our or many other samples solutions.

Let us open the previously saved TESTPROGRAM. Press the Load program button and
select "TESTPROGRAM" from the displayed program name list.
The program will be loaded and will look like the one below.

Note: Load program is only available when the IceCube has established communication with
the PC. Only then is Load program visible.

Our test-program is fairly small, so it will fit on one display screen. Following are the various program
steps.

Step 1 Step 2

Step 3

Step 1 lowers the temperature in the chamber from 20.00°C to 0.00°C with a slope of -2.00°C/min.

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Step 2 holds the chambertemperature at 0.00°C for 5.00 minutes.


Thereafter, step 3 lowers again the chamber temperature from 0.00°C to -40.00°C with a slope of -
1.00°C/min.

We assume that the crystallization temperature will be around -7 to -10°C. So we have covered with this
program the temperature range of interest.
At the moment, we see only the program temperature. It is the green curve or line.
When the program is running, we also see a red and blue curve. The red curve represents the chamber
temperature, whereas the blue one represents the sample temperature (if inserted into a sample). The
sample sensor is only monitoring the temperature behavior in the sample and does not take part in the
temperature regulation of the chamber. However, for other functions like Autoseeding, the sample sensor
can be selected to trigger Autoseeding. More on this subject in section "Autoseeding".

Before the start of the program, only the Chamber- and Sample Temperatures are displayed. As soon as
the program run is started, the Program temperature and the Time display will be shown as well.

Sensor 3 temp. and Sensor 4 temp. will be active only when additional sensors are installed. In the
standard configuration, the IceCube comes only with two sensors - Chamber and Sample sensor.

Inspect the level of Liquid Nitrogen in your supply vessel. Also check that the vessel is pressurized and
the liquid valve is opened. This freezing cycle might need anywhere from 5 to 10 liters of Liquid Nitrogen
(standard program).

Note: Use only supply vessels which are approved and confirm to the local regulations about those
pressure vessels. Only trained personnel is allowed to operate such vessels. All necessary and required
safety precautions must be followed. If in thought, please contact your safety adviser.

You can see that the "to Start Temp" has changed its color to blue. A sign, that it is ready
to be pressed.

If all conditions are right, press the "to Start Temp" button.
As soon as the "to Start Temp" is pressed, the IceCube will try to reach the start
temperature. At the same time, the button "to Start Temp" will change to "Start".

At this point, it is possible to start with the actual program run. However, in most cases,
we want to wait until the freezing chamber reaches the start temperature. When the start
temperature is reached, the Start button will be "illuminated".
This is the time to insert your samples into the freezing chamber.

Please note also the "lamps" in the Status Area. The Fan light as well as the Online (CTS)
light should be ON, telling us that

a) the fan is running and


b) communication (PC to Freezer) is established.

In addition, depending on the initial temperature of the freezing chamber,


the Cool and / or Heat lamps should be turned ON and OFF in a periodic manner.

As seen on the following screen, we have reached the start temperature, the "to Start Temp"
button has changed to Start and is illuminated.

Insert your sample solution / material, etc. and press Start.

Immediately, the IceCube starts to run the freezing process. The screen changes and
displays three curves. The green one represents the program-, the red one the chamber- and
the blue one the sample temperature.
Occasionally, if all temperatures are nearly the same, the three curves
will overlap and only one curve might be seen. If you have activated Actual Program

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position - show and display numerical temp. in the Display Settings menu, the program temperature will
be shown in a little window next to the temperature curve.

The freezing process will now proceed. As soon as Start has been pressed, the Start button changes
back to "to Start Temp" and will be disabled. Other buttons however will be activated.

As we can see in the next screen, the three


temperature curves are clearly visible. The
chamber (red) temperature is exactly
following the program (green) temperature
(overlapps). The sample (blue) temperature
is lagging behind, because the sample
needs more time to cool.
We are now waiting for the
moment at which the
sample temperature will be
rising very rapidly. This will
be our crystallization
temperature of this sample.

As it happens,
crystallization took place at a chamber
temperature of 11.9°C. At the same time,
the sample temperature was at -10.3°C. It is
the chamber temperature that we are
interested in. In addition, we will further
investigate the sample to see how long it will
remain at the elevated temperature. Small
samples will continue to cool faster than
larger samples. Actually, what you see
below the blue curve is the amount of latent
HEAT. It will be our task to modify the test
program in such a way as to compensate for
this amount of heat. Ideally, the blue curve
should remain parallel to the green / red
curve. However, this will not be possible. The phase
transition from liquid to ice will always be seen. It is
on us to find a curve that will minimize the increase
of heat. This procedure is called "optimization".
As we can see below, the sample curve tries to
reach the program curve again. This is then the
case when all the liquid has changed to solid (ice).
As long as there exists a mixture of liquid and ice,
the temperature of the sample remains nearly
constant.

For now, we have gathered enough information to


modify our test curve. Therefore, we will now stop
the freezing process. Press Stop. You will be asked
if you really want to stop? Press YES.

As soon as you say YES, the freezing process will be stopped and the total freezing process curve will be
displayed.

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The freezing process has been saved to disk automatically. There is no need for additional saving. Just
remember, the freezing process file will be found in YOUR specified data directory location under the
name convention (e.g.):

LAST_yyyy-mm-dd_hh-mm-ss.prc

Operation Panel

Seeding
Seeding - Manual - (works only if optional seeding unit is installed): Seeding is another way of
freezing a sample. Instead of using a heat compensation curve (will be shown later when optimizing a
curve), the main task here is to uniformly grow ice crystals around cells, embryos, etc. Please consult the
section on "Seeding". A program can be written such as to perform seeding automatically. Please read
the section on "Autoseeding".
For seeding, optional equipment is required.

Hold
Hold: During a freezing process, the operator has the possibility to manually suspend the run. This is
accomplished with the Hold function. When Hold is pressed, the current temperature will be held constant
for as long as the Continue button is pressed. The Hold button will be toggled to Continue after it is
pressed and vice versa.
In other words, a Hold / Continue operation adds time to the total length of the program. One application
would be that the operator needs more time to fill up the freezing chamber. So at start temperature, he
just presses Hold. When the chamber is loaded, he Continues with the freezing run.

Next
Next: The Next function performs the task of jumping to the beginning of the next step before the
previous step was finished. For example, if we are in the middle of program segment (step) 1 and we
would like to proceed rapidly to the beginning of step 2, we just hit Next. In the section "Optimizing a
Program Curve", we will discuss the usefulness of this function. Next will not work during a Hold.

Load program/Stop
Load program button shows up first to load an existing program. Once a program is selected (loaded),
this button changes to "Stop".

Stop: As the name already implies, Stop will stop the freezing run. But before this happens, you will be
asked to acknowledge this action.

You can then choose YES or NO.

The process file will be saved under the following file name convention.

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Sample- or Patient name_Date_Time.PRC


e.g.

Huber_2003-03-18_13-47-26.prc
(max. 8 characters)_year-month-day_hour-minutes-seconds.prc

Also, you will be asked if a standard report should be printed.

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Writing the final program - Optimization (as an example)


In this section, we are going to modify our test program such as to compensate for the heat of
crystallization. In addition, we will add the remaining steps to bring the sample down to a temperature of
about -120°C. At this temperature, most of the samples are then transferred to a long-term storage
vessel.

Let us recall what we have so far.

Step Temp - °C Slope - °C/min. Time - min.


0 20,00 0,00
1 0,00 -2,00 10,00
2 0,00 0,00 5,00
3 -40,00 -1,00 40,00

We also learned that the crystallization of the sample takes place at a sample temperature of
-10.3°C. The corresponding chamber temperature was at -11.9°C. Since we want to compensate the
increase of heat in the sample, we have to rapidly lower the temperature in the chamber just an instant
before the warming starts. So we will modify step 3 above to cool down rapidly from -11.4°C (about 0.5°C
less as the chamber temperature at crystallization) to -40°C with a speed (slope) of -60°C/min. From past
experiences, it was found that compensation temperatures of -40°C or -50°C are suitable for many
applications.
At -40°C, we will remain at this temperature for some period of time. Again, from experience, for vials we
will keep the temperature at -40°C for 0.5 minutes. Then, we will increase the temperature again to about
-25°C with a slope of +10°C/min. To slow down the heating process, we add another step with a smaller
slope. From -25°C we go to -17°C with +3°C/min. Then, we continue to cool with a slope of -1°C/min.
down to -30°C. At this point, the sample(s) should be all frozen. Now, we can speed up the cooling
process. From -30°C to -70°C we cool at a rate of -5°C/min. Thereafter, we approach our final end
temperature of -120°C with a cooling rate of -8°C/min.

In tabular form, it looks like this:

Step Temp - °C Slope - °C/min. Time - min.


0 +20,00 0,00
1 0,00 -2,00 10,00
2 0,00 0,00 5,00
3 -11,4 -1,00 11,40
4 -40,00 -60,00 0,48
5 -40,00 0,00 0,50
6 -25,00 +10,00 1,50
7 -17,00 +3,00 2,67
8 -30,00 -1,00 13,00
9 -70,00 -5,00 8,00
10 -120,00 -8,00 6,25

Use the same procedure as described above in the section "Writing a Test Program" to enter these
program steps. Call this program "FullProgram".

The result will be shown below.


In the display area, we see the graphical representation of the freezing process curve. On the right, we
have the corresponding tabular listing. It should be identical to the table we have worked out before.

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If not yet done, "save as" the new program under the name "FullProgram".

Prepare a new sample of the same material and quantity. Use the same type of vial. Vials from another
manufacturer might have different heat transfer coefficients. So please keep all parameters the same.
Also, do not use a pressure vessel with a different pressure. It will definitely change slightly the
temperature point at which crystallization takes place.
Just remember, this is our first real freezing process curve. In most cases, this curve has to be modified
repeatedly to get the optimum freezing process for our sample. According to the results of this freezing
run, we will have to further adjust (optimize) this program curve.

Check again if:

the pressure in the supply vessel is adequate,

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the liquid valve is open


new sample is on the sample sensor
the chamber is at about ambient temperature
the program "FullProgram" is loaded
the chamber lid is closed.

Press "to Start Temp".

You will now be asked to fill in some sample or patient information. It is mandatory to enter at least a
Sample / Patient name. It uses this info for creating a unique file name. Enter "Test" for Last name.

As a first step, the chamber will be brought rapidly to the start temperature. In our example, this is +20°C.
As soon as the start temperature is reached, the blue Start button will light up, signaling that
the freezing chamber has reached the start temperature and that the sample can be loaded
into the chamber.

In the screen below, we have on purpose, forgotten to press the Start button. Nothing much
really happened. The chamber temperature will remain at the start temperature as long as we do not
press Start or Stop or the Liquid Nitrogen runs short.
Here we can see how constant the start temperature remains. When Total View is selected, the freezing
process file (green) will also be displayed.

The system has stabilized at +20°C. Press Start to start the freezing process. It will now take about 26
minutes before we reach the crystallization temperature again.

Normally, it takes a few runs to get a good freezing process curve. Depending on the sample material and
its containment, optimizing a curve can be simple and quick or just be the opposite. We will see in about
26 minutes.

If we selected in Main Menu - Display - Settings "Actual program position" - display numerical temp., we
will also see the numerical display of the program temperature during the freezing run on the screen.

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While this program is running, let us look at


Autoseeding.

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Special Function - Autoseeding


As already mentioned before, there are some special functions built into the IceCube
program. One is the Seeding. Seeding can be programmed so that it is initialized
automatically.
We can find this function in the Main Menu - File - Freezing program - Edit existing or
Create new.

The function "Trigger" is not available on the 14S Freezer.

Setting the Autoseeding Level


At the beginning of this manual, we have followed the
Program path. Now, we will look into the Autoseeding
path. Click on Autoseeding. Following screen will
appears

For IceCubes without the Autoseeding option,


Autoseeding should be disabled.

If Autoseeding is installed, the operator has two possibilities to initialize the seeding.
by temperature of any sensor or
by time

If temperature is chosen as the seeding criteria, we have to tell the


system at what temperature to start and on which Sensor to measure.
Any temperature sensor can be selected. In general, the sample sensor
is used for monitoring. As an example, above, we have found
crystallization talking place at a sample temperature of 10.9°C. This was
measured with the sample sensor in the sample (vial). Seeding should
take place prior to the self-crystallization. So we choose a temperature
somewhat less than 10.9°C. For a first try, we should enter 10.4°C in the
temperature field and select the Chamber sensor as measuring criteria. Press Enter. As soon as the
Enter button is pressed, a horizontal line on the graphics display will indicate when Autoseeding will be
turned on.

In addition, we have to tell the system for how long the seeding valve should open - Active time. Past
experience has shown that seeding should not last longer than about 30 seconds. If it lasts longer, the
freezing behavior of the sample might change.

Note: In order to understand what seeding is doing, please refer to the section on "Seeding".

If crystallization of the sample is not initialized within 30 seconds, the sample


temperature is still too high (too warm). If the crystallization temperature was
determined accurately, seeding times of 10 seconds are not uncommon. The
shorter the seeding time, the better for the sample.
The second way to initialize seeding is on Time. Basically, if we run a
freezing process, the crystallization temperature will be reached in a certain
time. So this criterion can be taken as well. If we know that it takes 26.4 minutes for the sample to reach
10.4°C, we can enter in the time field 26.4 minutes. The valve open time should be the same as above.
No more than 30 seconds. The shorter, the better.

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Now, let's come back to our resulting freezing curve.

As we can see, the sample temperature is


still rising around the crystallization
temperature. So apparently, the quick
temperature compensation at -11,4°C was
initialized too late. So we have to modify
our current freezing program
"FullProgram".

Step number 3 has to be changed from -11,4°C to let's say 10,3°C as seen in
the tabular listing on the left.
Save the newly modified curve under the name "FullProgram2".

Again, we have to re-run the sample. As already mentioned earlier, always


start at the same conditions. For instance, freezing chamber at 20°C, sample
at 20°C, new sample of same material and quantity as in the previous run,
sample sensor inserted into the geometric center of the sample, same
pressure vessel, etc.
The result of this curve will let us know if we have to continue to optimize the
curve or if it is already acceptable. Normally, we would have to go through
another 2 to 3 runs before the curve "might" be optimized.

Without documenting here the same curves over and over again, it was found that when running the
program on the left, the sample temperature still increased at the crystallization temperature. So
therefore, Step 3 was changed to -9.5°C and the freezing process started again.
It will take about another 26 minutes before we can see the results.

We can see that even at -9.5°C, there is still an increase at the


sample temperature around crystallization. So we will once more,
modify Step 3 down from -9.5°C to -8.5°C and see what happens.

If we compare the current freezing process with the initial one, we


will discover that the area under the sample (blue) curve has
become smaller. This in turn tells us that the low temperature in the
chamber has compensated some of the heat given off by the sample
during the crystallization.
Having now changed step 3 to -8.0°C, we rerun the sample again,
assuming the system is back at starting condition.
The result of this run is visible. As you can see, the increase of heat
in the sample is already smaller than it was ever before. Also, the
temperature increase of the sample is already somewhat
compensated by the lower temperature in the chamber. However, we still have to compensate a little
earlier. This time, we set step 3 to -6.5°C and see what happens.

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Once we know the characteristic behavior of our material, we might


be able to speed up the optimization of our freezing processes.

Initially, the crystallization temperature of the 10% Glycerol / Water


solution was found to be at a chamber temperature of -10.9°C. Now
we are down at about -6.5°C. Naturally, the rapid cooling will
somehow change the cooling behavior of our sample. Since
different sample materials are behaving differently, we have to
follow this procedure by trial and error.

Now, the curve on the left


looks again better than the
previous one but still is not optimized. So we try once more to lower
the rapid cooling point (step 3) to -5.5°C. At the same time, we will
slightly change step 7. As it seems, once the sample temperature
increase ceases, the heating time or speed of step 7 (+3°C/min)
takes too long. So we increase the speed of heating to 5°C/min.

Here, we see that with step 3 set to -5.5°C, the rapid increase of the
sample temperature starts to flatten out. This is already a good sign
that we are very close to our goal - getting a curve that is as flat as
possible.

So again, once more we increase step 3 to -4.5°C.


Our tabular listing now looks like this:

Now, let us run the sample one more time and see what happens.

The final freezing process curve is shown below. As you can see
now on the enlargement, the increase of temperature in the sample
has almost ceased. As a comparison, look back at our initial freezing
process and compare the area below the sample curve. Can you see
the big difference? We just have optimized our freezing process
curve.

If you have enough time, you can play some more with the
temperature profile. Perhaps you get the curve even flatter.

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Below, the same curve as above is shown but only


the scaling factor has changed. Again, we can see
that the increase of heat in the sample is
minimized and fairly well compensated by the
temperature profile below.
We are now at the end of describing how to
optimize a temperature profile. It is now up to you
to either accept the program curve as is or to play
some more to make the curve even flatter.

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Setting the Restart Level


(only if no additional emergency power source / UPS is connected)

Another function in the Edit control window is Restart. After a power failure, the IceCube freezer tries to
re-establish the temperature conditions it had before the power failure occurred, depending on the
settings in Restart.

First, a critical temperature and time range can be set. In other words, we can
specify an area where our sample is most damageable. Naturally, this will be
around the point of crystallization.

So we will set the limits accordingly to our freezing process curve. Both criteria
have to be met; the temperature in the sample after power return has to be
within the temperature and time limit settings. In our example below, it will look
like this.

The temperature limits are set


approximately between 2,5°C and
-24°C. The time limits are set
approx. between 15.6 and 40
minutes. Let us analyze these
settings.

Case 1: Above or around zero


(0°C), the sample is not frozen yet.
Also, we have not reached 25
minutes yet. After power comes back, the IceCube will continue automatically with the freezing process.

Case 2: The sample temperature is below -24°C and the program run time is over 40 minutes. The
sample is practically frozen and the IceCube will automatically continue with the freezing process once
power is reestablished to the instrument.

Case 3: The sample temperature at the time of power return is within the upper and lower limit (between
2.5 and -24°C) and the power failure occurred within the left and right time limit. The freezing process will
not automatically continue. In this case, an alarm will be issued, signaling to the operator to take action.
The responsibility is now on the operator to decide what to do. If the sample has not started to crystallize
yet, it might be possible to continue with the program run. If the sample is nearly at the end of
crystallization, it might be possible to continue with the program run. However, if the sample is just at this
point where the heat of crystallization is removed, your sample might be spoiled. For important sample
material, we always recommend using an Uninterruptible Power Supply (UPS).

Also, we can set the Maximum non-critical power fail time to bridge the time required for emergency
power supplies to come up. Most hospitals will have such emergency power supplies. If not continuously
operated, it might take a few seconds to start such systems.
If power is interrupted, the IceCube system will stop working. However, if this interruption is only of short
duration (a few seconds), the IceCube will automatically continue with the freezing process as if nothing
would have happened.
The internal microprocessor of the IceCube stores all of the settings and temperature readings prior to the
power failure and can therefore immediately (within 1 sec.) continue with the program run after the power
is restored to the instrument.
The external PC will need some time to boot up again and it might take a minute or two to see the actual
freezer program on the screen again. During all this time, the freezer operates properly in the
background. In addition, no data will be lost.
It is up to the operator to find out how long the emergency power supply needs to be activated. Add a few
seconds and enter it into the Maximum non-critical power fail time. As a good guess, 5-10 seconds might
be sufficient.

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Now keep in mind, during the power failure, the


temperature of the chamber and sample will
have changed. Depending on the length of the
power failure, this temperature change can be
small or large. The Freezer has now to decide at
what temperature it should continue with the
freezing process. For this purpose, we have provided two choices.

Continue with the freezing process at the temperature of the sample -Sample Temperature.
As an example, the temperature of the sample before power fail was at -40°C. After a power fail
of 3 minutes, the temperature of the sample increased to -35°C. When power is re-established,
the freezing process will continue with the program run at -35°C up to the last step of the
program. This type of setting will provide a smooth temperature change between the actual
sample temperature and the freezing process temperature.
Move to a preset chamber temperature and continue with the freezing process from there. Some
applications might require bringing the sample, after a power failure, to a preset temperature. In
this case, use the function preset temperature of xx.xx °C.

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Seeding
(Inducing Ice Formation under Controlled Conditions)

Auto(matic)Seeding
(Available as an option)
Auto-seeding or Automatic seeding allows the user to select a pre-defined temperature point at which
crystallization should begin. Setting a "seed" does this. To be more precise, one point on the outside wall of
the sample container (straw, bag or vial) is brought to a very deep temperature. The temperature of the
medium inside the sample container will also drop rapidly at this point. The temperature of the medium next
to this cool spot will have decreased down so far as to induce the crystallization process. The result will be
the uniform growing of ice crystals. Most important is the fact that the speed of this uniform crystallization
can be controlled. This means that it is possible to control the amount of heat given up within a period of
time, giving the system enough time to dissipate the latent heat of crystallization. If the temperature of the
sample material or sample solution is far (a few degrees) from the temperature of self-crystallization,
performing Autoseeding will result in a slow freezing process. As closer the temperature of the sample is to
the point of self-crystallization, as faster the sample will freeze when Autoseeding is turned on, thus being
able to control the amount of heat dissipated in time.

In addition to Auto(matic)seeding, seeding can be done manually as well. Manual seeding is described in
this manual under "Special Functions".

Below, you can see a typical auto-seeding rack for straws.

Note: An Instruction Manual for High Security Straws (CBS) can be found as a Supplement in
the Appendix.

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Maintenance
Leak Check
Check occasionally the connections of the Liquid Nitrogen Supply hose for leaks. Due to the high
temperature differences in the hose, shrinkage and expansion of the material might cause the hose to
become leaky. Just retighten slightly to fix this problem.
If the supply system is still leaking, check both sides of the supply hose as well as the connections on freezer
and supply vessel. Any damaged parts must be replaced by authorized personnel.

Warning: Do not turn the system on if the instrument or parts of it are


damaged.

Fog (condensed moisture) on the supply tubing and its connections will sink to the ground. This is normal
and does not represent a leak in the system. If no liquid is expelled by the supply tube, the system has no
leak,

Cleaning of the Freezing Chamber and Instrument


Caution: Before doing any cleaning work on the freezer, disconnect it from the power line.

Due to technical reasons, the freezing chamber can not be removed from the freezer housing. However, the
freezing chamber is fairly large and cleaning should not create any difficulties. Use only mild and soapy non-
scratching detergents on a wet cloth. The materials used inside the chamber are Stainless Steel, Glass and
Glass fiber reinforced plastics. Use only disinfectants which will not harm those materials.
After cleaning and disinfecting, wipe the chamber dry with soft cloth or paper.
The freezer housing itself must be cleaned only with a soft wet cloth.

Handling of Temperature Sensors


The temperature sensors should be handled with special care. Never apply any force on the sensors. Never
try to pull out a sensor from a frozen sample. Wait until the sample reference has changed to liquid again.
The sample sensor feed through is marked as blue. The chamber sensor is marked as red. Never try to
insert the chamber sensor (red) into a sample. Unknown chamber temperature behavior will be the result.
Use only the sample sensor to monitor the sample material.
Do not bend or drop a temperature sensor. Limited warranty conditions apply.

Cleaning

Caution: Before doing any cleaning work on the freezer, disconnect it from the
power line.

Due to technical reasons, the freezing chamber cannot be removed from the freezer housing. However, the
freezing chamber is fairly large and cleaning should not create any difficulties. Use only mild and soapy non-
scratching detergents on a wet cloth. The materials used inside the chamber are Stainless Steel, Glass and
Glass fiber reinforced plastics. Use only disinfectants which will not harm those materials.
After cleaning and disinfecting, wipe the chamber dry with soft cloth or paper.
The freezer housing itself must be cleaned only with a soft wet cloth and a mild detergent. Parts made of
Stainless Steel must be treated similarly.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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Settings on the PC

Disabling of Start-Up Error Message


(Should be done by the system administrator. SY-LAB will not take any responsibility for changes made in
the BIOS, Registry or any other software related items. The description below can vary depending on the
type of PC and operating system used. Therefore, we see the descriptions below only as a hint to help you
along).

Computers that are delivered from SY-LAB together with an IceCube are pre-configured. However, if you
supply your own PC or the original PC needs to be changed, some settings have to be changed.
In normal operation with the Freezer, no keyboard is connected to the PC. Therefore, the computer has to
start without a keyboard. In order to do so, we have to disable in the System-BIOS the entry

"Hold on Keyboard Error"

On some computers, this function cannot be disabled in the BIOS. A keyboard error will be displayed on the
screen "press F1 to continue". Ignore this message, the computer will still boot up after a few seconds.
Note: For BIOS changes, a keyboard has to be connected to the PC.

If this is different on your computer, please refer to the PC's User Manual.

Disabling Request for Ctrl - Alt - Delete under WIN 2000


under WINDOWS® 2000

For automatic Freezer restart after a power-failure, the login request Ctrl-Alt-Delete has to be disabled.
Press Ctrl-Alt_Delete to begin
Ctrl-Alt-Delete helps keep your password secure ----Ctrl-Alt-Del

User name: Administrator


Password: ...................... (leave blank)
Log on to: ...................... (leave blank)
□ log on using dial-up connection

OK Cancel Shutdown Option <<

To remove the above login request, boot up the computer with mouse and keyboard connected.
Depending on your Windows® operating system, here we show it for Windows® 2000, go into

Settings - Control Panel - Users and Passwords - Advanced - Secure Boot Setting
□ Require users to press Ctrl-Alt-Delete before logging on

Remove the checkmark () from the checkbox and restart you computer again. This time, you will not be
asked for Ctrl-Alt Delete anymore.

Please Note: Not all computers feature an automatic restart after power loss. For such systems, the PC
System must be turned on manually. However, no data will be lost since all data is saved internally in the
IceCube and are transferred to the PC when power is re-established.
However, you will still be asked for a User and Password. Since you did not enter a Password previously,
there is no need to enter one. The User name will be already assigned. However, at this point, even though
no keyboard is necessary, you still have to press on OK. This creates a problem for the automatic restart
feature of the IceCube.

Therefore, the Auto Log On feature has to be enabled. Since this procedure will be different among
computers, we suggest leaving this up to your PC's system advisor. It involves changes in the PC'S registry.
Caution:

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Again, we want to point out that the settings should be done only by your PC Advisor. Any wrong changes
made to the registry could lead to total failure of the PC system. Make a backup of your registry before
making any changes to it.

Start - Run - <regedit> (Enter)

You are now in the REGISTRY

Go to: HKEY_LOCAL_MACHINE \ SOFTWARE \ Microsoft \


WindowsNT \ CurrentVersion \ Winlogon

following should be entered:

AutoAdminLogon (REG_SZ) = 1 (0 = No automatic Log.on)


DefaultDomaineName (REG_SZ) = <Your Domaenname>
DefaultUserName (REG:SZ) = <Your Username>
DefaultPassword(REG_SZ) = <Your User Password> (if one was specified earlier, otherwise, this field will
be empty).

Exit.

Disabling Power Save Options


All power save options (features) must be disabled. These are the Screensaver, no Power down, no Standby
and no Hibernation.

The disabling procedures vary according from operating system to operator system. Here, we give some
examples on how to disable these functions on our currently available XP Professional operating system.
Consult the operating system manual for your system.

Windows® XP-Professional
Press the right mouse button while pointing into a free area on the Desktop.
Choose “Properties”. - “Screen Saver” and set the “Screen Saver” field to None.

Then,
Click on the button “Power …..” and set all “Settings for ….” to “NEVER”.
e.g. Turn off monitor: Never
Turn off hard disk: Never

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System standby: Never


System hibernates: Never
and acknowledge with OK.

While in the “Display Properties” screen, select “Hibernate” and remove the mark from
“Enable Hibernate Support”.

Now all power save options, etc. should be disabled.

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Freezer Software (Re-) Installation


Freezer Software (Re-) Installation
(PC knowledge required).
Note: Please read all the applicable license agreements. Install only programs for which you are agreeing to
these agreements. If you do not agree, do not continue to set up that particular program. Preinstalled
programs must only be used when you are fully agreeing to the various program license modules.
............................................................................................................................................
IceCube Series - License Agreement
The product IceCube Series (synonyms "program" or "software") is the intellectual property of SY-LAB
Geräte GmbH, Austria.
It is forbidden to publish, recompile, change or copy the program or parts of it. Diagrams or results obtained
with the IceCube that are the property of the program owner are accepted. For the reason of data safety it is
allowed to make one copy of the program CD-ROM.

To use the IceCube program, a license from SY-LAB is necessary. It is not mandatory to have an IceCube
instrument to obtain a software license. A license is granted as a single place license in form of a hardlock
that is necessary to operate the IceCube Series instruments.
The loss of this hardlock is therefore equivalent to the loss of the license. If only the software CD-ROM gets
lost, the license is still valid and a new CD-ROM can be ordered without additional license fees.

Free updates are included in the license as long they are aimed to fix possible bugs. Program extensions are
not free and are not included in the original license

SY-LAB disclaims liability for any loss in connection with direct or indirect usage of the program. The users
by themselves are responsible for data safety and data protection

COPYRIGHT:
----------

MS Windows 95/98/NT/2000/XP are protected by Microsoft Corp. USA. IceCube (c) 2004 SY-LAB GmbH.
Austria.
All rights reserved

Purkersdorf, 27.5.2004

--------------------------------------------------------------------------------------------------------------

Every IceCube Freezer will be accompanied by a Program CD,


a calibration diskette and a hardlock.

Freezers which are delivered with a PC from SY-LAB have


already the software preinstalled on the PC. Just connect the
hard-lock on one of the USB ports, make all hardware
connections and start the IceCube Series program from the
desktop.

Note: To run a “validation” IceCube Series Freezer program,


an IceCubeVal hardlock has to be installed. It will not run with
a standard hardlock.

On PC's not supplied by SY-LAB, the software has to be


installed as follows.

Note: The following freezer software installation assumes that you have a correctly working PC system with
Windows 2000 or XPP pre-installed. SY-LAB will not be responsible for any malfunction if other operating
systems are used.

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On systems with Touch Monitor, a keyboard and a mouse are needed during the software installation.

1. Insert the IceCube Series Software CD into your CD-ROM drive. Setup program should start
automatically. If not, start the setup by selecting the file “Install.bat” on the IceCube Series Software
CD.
Note: Do not connect the USB Hardlock and the USB connection from the touch screen at this point.

The Hard lock installation program will be shown on the screen.

2. The TouchScreen Software will now be installed. You will now be asked to set the

Destination Location (Next); Select Start Menu Folder (Next),

Install (Installing) and Finish (Finish) the setup procedure.

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3. Now, the IceCube Series Program welcome screen appears. Press Next.
Agree to the License Agreement (Yes), choose the Destination Folder (Next) and choose the
Program Folder (Next).

The Setup files will be decompressing, icons created and the setup completed. Close the setup screen and
press Finish.

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The IceCube Series Icon will now be shown on the Desktop screen.

4. At this point, it is advisable to re-start you computer system. During the re-start, the error message

“Error 7: Hardlock not found” will be displayed. Ignore this error message.

5. Now, plug the USB Hardlock into one of the available USB Ports and wait until the device is installed
and ready. Unplug the hardlock and do the same on all other USB ports. The hardlock device is then
known at all USB ports and it does not matter at which USB port the hardlock is connected in the
future. Finally, leave the Hardlock connected at any USB port.
6. Attach now the USB TouchScreen cable to any USB port.
The welcome screen appears. Choose “No, not this time” and press Next.
Also, select “Install Software automatically”.

7. The Windows Logo test will not be passed. Ignore this message and select “Continue anyway”.

The TouchScreen software will now be installed automatically. At the end, press Finish.

Follow the instructions in the next section.

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TouchScreen Calibration
Before the first use, the TouchScreen monitor must be calibrated.

Go to START – All Programs – Touchscreen V1.00-USB… - Touchscreen Control Panel V1.00-USB.


The TouchScreen Control Panel appears on the screen.

Select the TouchScreen device “Device #2048…….” And press OK.

The TouchScreen Control Panel will open.

Set all entries to the ones shown above (if not already in this configuration).

Select “Calibration”.

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The Calibration Screen opens.

Calibration
Spot

Starting from the top, press as exactly as possible on the center of the Cross Mark. Then, the next Cross
Mark will be show. Do the same there until the “Calibration is complete” screen shows up.

Try now to touch a few points on the screen and check if the screen cursor moves to the new location. If yes,
close the window by pressing Yes. Also, EXIT the TouchScreen Control Panel.

Note: In case of a power failure, the IceCube Series program should start up by itself. For this to happen,
place the IceCube software icon into the “Auto-Startup”.

Now, the IceCube software can be started by pressing onto the IceCube Series Software Icon on the
desktop. The welcome screen appears and you are asked to enter your User Name and password. More on
this in the section “Program Log-In”.

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ImageLoader (Subsystem Update)


Concurrently to the IceCube program installation, an ImageLoader program was installed.
This program will allow you to update the Freezers internal operating program. If such an
update becomes necessary, SY-LAB or your representative will provide you with a small
xxx_yy.IMG file either on diskette or per e-mail.

To upload this upgrade file onto the freezer, start the ImageLoader program.
(found in the folder C:\Program Files\SY-LAB\IceCubeSeries)
Following screen will be displayed.

Select the *.IMG file (in our example - P18_01.IMG) and turn the Freezer ON. If the freezer was already
turned ON, turn it OFF for 2 seconds and ON again.
The upgrade file will be uploaded to the freezer. A progress bar, located right under the file name, will
advance from the left to the right, indicating the upload progress.

The new version of the freezer operating program can then be checked in the Freezer program under Main
Menu - Version Info as seen below as an example.

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Inserting Your Own LOGO


The IceCube software allows you to insert your own company or institute logo. Some
space has been reserved for this on the lower right of the screen, just above the SY-
LAB Logo. You can only add a logo once. In addition, the SY-LAB Logo is not
replaceable

The size of the logo should be 106 by 122 pixels in size in bitmap (*.bmp) format.

With no prior logo installed, double press on the dark area


above the SY-LAB logo. A file selection screen opens. Enter
the name of the logo file you would like to insert and press
OPEN. The logo will be placed into the empty black area like
shown below.

As long as you do not exit the Freezer program, you can replace the logo and play
around until the appearance suits you.
As soon as you exit the freezer program, the logo file will be permanently displayed.

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Troubleshooting
Problem Diagnostic / Solution
No Power to the system Check if there is power in the wall socket. Use a
Voltmeter to determine the correct voltage
output. Compare the voltage reading with the
voltage shown on the freezers type plate.
Are all components, printer, monitor freezer and
PC connected to a power distribution box?
Check!
Is the power distribution box plugged into the
wall socket?
Are all power switches of the single components
turned on?
Power to some components but not Check the single components for:
all Power rating (voltage), power connection, broken fuse,
power switches turned on?
After turn on, the PC gives error Enter your PC-BIOS and de-select this function. Check
message "Hold on all errors - No your PC's user manual on how to do this.
keyboard connected"

Power to PC, Monitor, Printer, and Message "Error 7: Hardlock not found" shown on
Freezer box ok. screen. The IceCube Freezer Series software is
IceCube software does not start. copy protected and needs a "Hardlock" to
function. Make sure the hardlock is installed
either on the parallel port (older version) or on
one of the USB-ports (newer version). Consult
the section on "Freezer Software Installation"
Windows Screen comes up normally but freezer
program still does not start.
Program fault - re-install the freezer software
Freezer-Icon on desktop points to a wrong path
Program Log-In not successful 1 Wrong "User" or "Password". First time users
can log-in using for both entries "sylab". The
Freezer Administrator should create a new User
Name List (also for him) with or without limited
access to the system. The factory set Log-In
name should then be deleted. See section on
"User Administration"
Program Log-In not successful 2 For first log-in, use sylab for UserName and
Password. In addition, the password entry is
case sensitive (distinguished between upper and
lower case letters). The UserName is not case
sensitive.
Once individual users are defined, each user has
Password lost / forgotten to Log-In using his specific UserName and
Password.
In case of a lost / forgotten Password, the
System Administrator having Administrator rights
can re-assign a UserName and Log-In
Password. The Password can then be changed
again by the individual user. Thereafter, the
Administrator has no way of knowing the users
individual password.
Note: If the Administrator also forgets his

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Password, the IceCube Freezer Series Software


has to be re-installed. Therefore, it is advisable
that two independent persons know the
Administrator Password.
Power to PC, Monitor and Printer The freezer electronic (box) is either not turned on, the
ok. communication cable to the PC not connected or the
IceCube software starts ok. freezer box fuse damaged.
"Online (CTS)" LED is OFF. Check all of the above, replace fuse if needed.

Check also the COM port settings in the freezer program


- Main - Maintenance - Communication and choose the
correct COM Port.

Note: When using an USB to RS-232 Adaptor (not part


of SY-LAB specification), it will be necessary to set the
Adaptor (in the Device Manager) to work on COM1 to
COM4 only, besides installing the necessary drivers.
After power failure, the IceCube Place the shortcut icon in the desktops "Start-up"
Program does not start menu.
automatically Also, deselect in the BIOS (not everywhere
available) the request for a keyboard except on
14S-A, where a keyboard is needed.
Disable the request for Ctrl-Alt-Del (Win2000).
See section on "Settings on PC"
Printer seems not to work After issuing a print command, it might take up to one
minute before the printer produces a print-out. Do not
touch any printer buttons during this period of time.
Other reasons, please check:
Power connection (power supply)
Printer turned off or not connected
Printer out of paper
Ink cartridge empty
Paper jam
(consult the Printers Operator Manual for further info)
Monitor shows vertical lines after Sequential turn-on procedure was not followed.
turn on Synchronization-problem.
Unplug and reconnect the power plug from the monitor.
This will re-synchronize the monitor and should cure the
problem.
Touch Monitor not working Assuming the correct software setup of the Touch
monitor as described in the user manual, check if the
USB connection is properly seated. Unplug shortly the
USB cable. Reapply and see if the problem has
vanished. Further troubleshooting on PC-System level
(see PC Handbook).
No signal to Monitor VGA Monitor cable not connected to either the PC or the
Monitor.
Chamber does not get cold during No pressure or no LIN in supply vessel.
cooling cycle Liquid valve on supply vessel turned off.
No supply tube connected between freezer and
supply vessel.
Magnetic valve does not open.
Reasons: Moisture caused MV to get stuck, dirt in
system, no electrical signal to valve. Let freezer warm
up, try again. If MV still does not operate, an electrical
problem might be the cause.
Supply tube connected to gas-phase of supply

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

vessel instead of to the liquid phase.


Chamber gets cold with no Magnetic valve slightly leaking - Listen into the
program running chamber compartment. A slight "zzschsch" can
be heard. Either the valve seat is dirty or
defective. Clean or replace (call service
representative)
Magnetic valve badly leaking - Moisture (Ice) or
dirt in the system. Magnetic valve stays open
due to one of the above.
Replace or clean (call service representative)
Freezing curve oscillates too much Check the supply tube and the pressure vessel
(+/- 3°C) for correct pressure. 0.5 to 0.9 bar. If pressure is
too low or too high, oscillation can occur.
Operating conditions not ideally (e.g. supply tube
to long or non-standard, supply pressure out of
specification, non-standard chamber furniture,
etc.
Chamber and / or Sample Sensor Various conditions could cause this effect:
Temperature do not follow the no or little pressure in supply vessel (out of
Program Curve of the Demo specification)
Program supply tube connected to the gas-phase of the
vessel
not enough LIN in the supply vessel
leak in the LIN supply line
blocked supply line (dirt or ice)
supply tube too long (non-standard)
temperature sensors defective, dirty or moist
chamber sensor inserted into a sample. The
chamber sensor must never be inserted into
anything. It is solely used for regulating the
temperature in the freezing chamber
Fan, Magnetic Valve, Heater not Chamber lid not closed,
working If physically closed but still get message, then
the electronic does not recognize the lid-switch.
Contact a SYLAB representative.
Heater not working Heater element broken
(run any program which has its Over temperature protection switch open or
start temperature at +40°C) defective
Contact a SYLAB representative
Wrong temperature display Bad contact on Pt-100 sockets. Check. One of
the Pt-100`s might be defect. The resistance of a
Pt-100 at about ambient temp. should be around
100 Ohms. Exchange if necessary. The sensors
can be switched among each other in order to
determine if the fault is on the sensor or the
electronic. Thereafter, an Ice-water calibration
becomes necessary.
The chamber is hotter than about 40 to 50°C.
The chamber is protected by a temperature
switch against over-heating. The heater circuit
will be turned off once a temperature of about 40
to 50°C is reached in the chamber. The
temperature switch will turn on again once the
chamber temperature comes down below 35°C.
Instrument does not cool Not enough LIN in supply vessel.
Not enough pressure in supply vessel (0,5 to 0,9
bar) - (7,5 to 13,5 psi)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Check if the gas phase valve is closed and that


there are no other leaks on the vessel
Supply valve on vessel is closed
Magnetic Valve does not open. Either it is frozen
or there is dirt in the system. In either case, the
valve must be removed, cleaned or be
exchanged. Should be done only through
authorized personnel. Ice particle or dirt might be
in the supply vessel. In this case, supply vessel
must be exchanged or cleaned.
Moisture in the system. Never, ever disconnect a
cold tube from the system. Moisture from the
surrounding air will enter the tube and condense.
When warmed up, water will then be carried to
the magnetic valve and will lead to malfunction
of the magnetic valve
Instrument does not heat Thermo switch open and/or defective
Heater element broken.
Contact SYLAB representative.
Log-In not possible For first log-in, use sylab for UserName and
Password. In addition, the password entry is
case sensitive (distinguished between upper and
lower case letters). The UserName is not case
sensitive.
Once individual users are defined, each user has
Password lost / forgotten to Log-In using his specific UserName and
Password.
In case of a lost / forgotten Password, the
System Administrator having Administrator rights
can re-assign a UserName and Log-In
Password. The Password can then be changed
again by the individual user. Thereafter, the
Administrator has no way of knowing the users
individual password.
Note: If the Administrator also forgets his
Password, the IceCube Freezer Series Software
has to be re-installed. Therefore, it is advisable
that two independent persons know the
Administrator Password.
Pressure display on Monitor (not Liquid phase valve on Vessel is closed
on 14S) shows different value as During opening and closing of the magnetic
the pressure display on the supply valve (it is normal)
vessel (+/- 10%) Either the pressure sensor in the freezer or the
one on the vessel are at fault.
Note: The pressure reading does not have to be
very exact. It is more like informing the user that
there is pressure applied to the freezer or not.
Temperature Sensor shows erratic Spikes on a temperature curve indicate a broken
values. Spikes on a temperature temperature sensor. In order to be sure if the
curve. temperature sensor or the associated electronic
circuit is at fault, exchange the chamber and the
sample sensor. If the problem shifts, the
temperature sensor is at fault. Exchange the
faulty sensor and perform an Ice-Water
Calibration.
Otherwise, there is some electronic problem in
the freezer electronic box. Contact a Sylab

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 94 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

representative.
"Seeding" does not respond Seeding option is not installed. Can be up-
graded in the field - Ask for quote.
Seeding valve defective, frozen or dirty - Call
Service
No signal to seeding valve - Call Service
Access to Seeding not granted (User Level)
"Trigger" does not respond (not on Trigger not installed
14S Series) Trigger is malfunctioning - Call Service
Access to Trigger not granted (User Level)
No Alarm output (for all alarms) Alarm(s) turned off - Check Main Menu - Options
- Device Settings. Each alarm can be turned on
and off. This applies also to optional external
alarms.
Internal Buzzer defective. Call Service
"Start" button does not light up Chamber temperature is oscillating too much
around the start temperature. See above in this
list.
Device Settings for "Start Temperature reached
condition" too narrow or time range to long.
Default: +/- 0,5°C for 20 seconds.
LIN Leak Due to high temperature deviations in the supply tubing
and its connections, it will become necessary to check
and /or re-tighten both ends of the supply tubing a few
minutes after a freezing run has started (when tube is
cold).

According to our own findings and the customer's feedback, the troubleshooting list will frequently be
updated.

Check at: www.sylab.at

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 95 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Freezer and Accessories

Furniture
No. Description Description 2
13-158500 Vial-rack for 1.8, 2 u. 5 ml f. 96 pos., made of CrNi
13-158570 Spacer complete
13-158510 Rack f. Straws 0,25 and 0,5 ml 36 pcs.
13-158515 Straw Distr. Block for 36 straws
13-158550 Autoseeding rack f. straws, 0.25 ml 43 pcs.
13-158551 Autoseeding rack f. straws, 0,50 ml 35 pcs.
13-158560 Exchange Magazin f. straws 0,25 ml
13-158561 Exchange Magazin f. straws 0,5 ml
13-158555 Autoseeding rack f. vials 28 pcs.
13-158705 Autoseeding Rack f. vials, vertical 28 pcs.
13-158552 Autoseeding Rack f. Straws. CBS 22 pcs.
13-158580 Exchange Magazin f. CBS Straws
13-158600 Rack f. Bloodbags, metal cassettes 11 positions
13-158610 Rack f. Bloodbags, univ. horizontal 6 positions
13-158650 Rack f. Cryo Sink
13-158755 Seeding manual, Topplate w. 16 Plugs
13-158750 Click Holder f. Straws w. Turnlock 64
13-158620 Rack vertical 6 pos.

SY-Lab reserves the right to change technical specifications without any prior notification.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 96 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Spare Parts 14S


Pcs. Description Part No.

Option Temperature Sensor 65 x 1.0 mm 13-156880


2 Temperature Sensor 65 x 1.5 mm *13-156885
Option Temperature Sensor 100 x 1.5 mm 13-156886
Option Temperature Sensor 160 x 1.5 mm 13-156887
Option Temperature Sensor 200 x 1.5 mm 13-156888

Option Flexible Stainless Steel hose, 1m 13-159010


Option Flexible Stainless Steel hose, 1.3m 13-159013
1 Fan-Motor *13-155400
1 Motor-Axis L43 *13-155411
1 Fan *13-154800
1 Seal for Freezing Compartment S-APF *13-153435
Option Touch Monitor, (Variant B,C,D) *13-156903

1 Magnetic Valve 12V (Note: 1 additional if Autoseeding is *13-155100


installed)
1 Safety Valve 1/4", 50 PSI *13-155080
1 Electronic Box, compl. *13-156000
Option Accessory package - up to 100 Liter Supply Vessel 13-159000
Option Accessory package - over 100 Liter Supply Vessel 13-159100
Option Fuse, 20x5/T 10A, 10 pcs./ pack (for 110V Units) 10-165039-01
Option Fuse, 20x5/T 5A, 10 pcs./ pack (for 230V Units) 10-165043-01
Option Additional 3 potential free Alarmrelais 13-159520
rd
Option 3 Temperature Sensor 13-159500
rd
Option 3 and 4rth Temperature Sensors 13-159510
Option Coupling in chamber for removable temperature sensor 13-159511
Option Update kit for Autoseeding - Vials and Straws 13-159532
Option Pressure Sensor Update kit 13-159540

*) Included in the standard IceCube14S shipment.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 97 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Appendix

Statement of Conformity (220 - 240 V AC)


(will be attached to instrument)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 98 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Statement of Conformity (110 - 120 V AC)


(will be attached to instrument)

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 99 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Block Diagram – Software

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 100 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Instruction Manual – Supplement

Autoseeding Unit for Welded High Security Straws (CBS®)

Supplement – Autoseeding Unit for Welded Straws (CBS)


SY-LAB Geräte GmbH., Tullnerbachstrasse 61-65, A 3011 Neupurkersdorf, Austria
Tel +43 2231 622 52 –0, FAX +43 2231 621 93
Internet: http://www.sylab.com/, E-mail: sales@sylab.com, ©2000-2004 SY-LAB Geraete GmbH.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 101 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Introduction
New Seeding Techniques
New seeding techniques have made it necessary to re-design the current Autoseeding system. Previous
systems used straws that were sealed at one end with either powder material or with sealing pins (balls,
etc.).
®
Nowadays, many applications require straws to be welded on both sides to prevent contamination (CBS –
High Security Straws).

®
Above, three standard straws and one CBS straw are shown.
The standard SY-LAB Autoseeding magazine could accommodate non-welded straws of size 0,5 ml and
0,25 ml.
®
The new system for CBS straws will be for straws of size 0,3 to 0,5 ml which are currently available (other
sizes upon request).

In order to use Autoseeding in any SY-LAB IceCube


freezer, the Autoseeding option has to be installed. This
can easily be determined by checking if an Autoseeding
connection is available. On the 14S and 15M/L freezers,
the connector is located at the upper right backside part
of the freezing chamber. A black rubber access cover
indicates that Autoseeding is not installed. Please contact
your Sy-Lab agent for more information.

In the following section, the handling of the Autoseeding


®
system, using CBS straws will be explained.

Note: The photos shown are that of a 14S Seeding System. However, this description also applies the 15M/L
IceCube freezer systems.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

On the right, a complete


Autoseeding system is
shown.

It consists of the seeding


base, straw magazine with
metal clamp, inlet and out-
let tubing as well as the
flow-through section.

Both, the flow-through


as well as the straw
magazine can be re-
positioned to hold different
length of straws. Also, by
re-locating the flow-
through section, the point Outlet
of seeding initialization with
can be set arbitrarily as Flow- locking
well. Through Status Area plate
Inlet

Loading the CBS® Straws


Position the straws on the
straw magazine as shown Envelope
on the right. Make sure
they are all aligned. Also,
the position of the straw
label (envelope) should be
such as not to touch the
flow – through section. Slot

Place the Clamp on the


straw magazine and make Slot
sure it sits well in the slot
(1) on both sides of the Clamp
straw magazine.
Flow-
Through

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
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SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Then, turn the clamp


upward (2).
The two pins at the bottom
of the clamp will snap 2
into its mating part as
shown below (3). 1

Pin

3
Mating
part (bottom
slot)
®
The CBS straws will be held in
position between the clamp
(Silicon sleeve) and the straw
magazine. Also, the straw
magazine is built in a way as to
exert a small force on the flow-
through. This way, the straw is
making a firm contact with the
flow-through system.
In addition, when turning the
straw magazine by 180°, the
clamp can be used to exert an
additional force onto the straws
to put even more pressure onto
the seeding point if necessary.

As can be seen on the left, the straw magazine was just


turned around to allow for the clamp to lie on the straws. The
point of contact between the straw and the flow-trough
section is such amplified.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 104 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

For a more comfortable


handling, the complete
straw holder (magazine and
clamp) can be removed
from the autoseeding base
(e.g. for transferring into a
storage container, etc.).

Positio
n of
Autoseeding Unit in Freezing Chamber
The Autoseeding Unit must be positioned in the freezing chamber such as to keep the air (Nitrogen) flow
unobstructed.
The base of the Autoseeding Unit has two open ends.
Place the Autoseeding Unit with the open end towards the fan (ventilator).

If this is not followed, the


circulation of nitrogen is not
adequate enough to cool the
straws down equally well.

The open tube end must be


connected to the LIN- Inlet LN2-Inlet Open end
port. of base-
The other tube end with the ment
locking plate must be fixed
in the exhaust port.
Please make sure the tubes LN2-outlet
are not blocked or jammed.

When looking through the


exhaust port, a little groove
can be seen. Just position

the locking plate such as to snap into place.

In this way, any liquid nitrogen that enters the flow through section will be
diverted out of the freezing compartment, thus, not altering much the
surrounding temperature of the straws.

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 105 / 106
SY-LAB Instruments GmbH. Austria Operator Manual - IceCube 14S

Part Numbers to Order


(up to 22 Straws)

Part Number Description Description 2


®
13-158552 Autoseeding System f. CBS complete High Security Straws 0,3-0,5 ml
®
13-158580 Magazine + Clamp for CBS --------------------“------------------
®
13-158581 Magazine for CBS (only) --------------------“------------------

12/05 JE

Operation Manual IceCube 14S – Ver. 1.21b Rev 003 © 2007 SY-LAB Geräte GmbH.
All Rights Reserved 2008-01-23
Page 106 / 106