AUTOMATION DIFFERENCES BETWEEN SEGMENTED & FLOW INJECTION
Four different approaches: ANALYSIS
1. continuous flow analysis
- segmented stream
- flow injection
2. discrete analysis
3. centrifugal analysis
4. dry slide technology
5. reagent package technology
CONTINUOUS FLOW ANALYSIS
- 1950: 1st developed by Leonard Skeggs
- Described as the way on w/c chemical & sample travels
through the instrument
- A type of sequential analysis w/c all samples pass through
the same continuous stream and undergo the same
analytical process at the same time
- All the assays in the profile were performed whether or
not they had been requested in the specific patient
- Unique characteristic: use of air bubbles in the sample &
reagent streams
CFA SEGMENTED STREAM
Principle:
- air injected into each stream as a source of air bubbles
w/c travel along the reaction system
- air bubbles: minimizes diffusion of reagents & mixing
between samples preserve integrity of each individual
reaction (stream segmented by air bubbles promote
sample integrity)
 takes measurements in a stationary flow all through w/c
the reaction stream passes
 current analyzers – analyzes 20 or more different
constituents in a single sample in less than 1 min
 # & spacing of bubbles are controlled & the
microprocessor enables the readings to be taken
between bubbles
 Single or multichannel
 Sample(HCl)mixing coil dialyzer (cresolphthalein
complexone)(diethylamine)mixing coil colorimeter
data processing
Typical application (serum Ca assay):
1. Ca (bound to CHON) + HCl CHON + unbound Ca-
2. Ca + cresolphthaleine complexone  complex product
3. Colored product + DEA intensified color
*read at 580nm to quantitate the amt of color & indicate level
of Ca
FLOW INJECTION ANALYSIS
Principle:
- direct injection of sample into a very small diameter
tubing was utilized for reactions & measurements
1. Reagent stream – pumped through a small tubing (0.5mm
diameter)
2. Sample slug is introduced directly into the stream allowing
immediate mixing a few seconds
- sample vol = 50ul or less; reaction time: short, often
less than 1 min
3. Mixture passes through detector (microprocessor)
4. Absorbance measured concentration determined
*fluorocarbon coats the inside of both samples & reagent
transfer probes. Thus eliminates contact between samples/
rgts & the inside wall of the probe
* fluorocarbon coats the sample completely forming a “shrug”
w/c is dispersed during air pressure
- Stored in ref/ freezer (depending on test)
- Microslide technology design: deliver the highest Px
1. elimination of air bubbles due to the design of the
flow tubing
- extremely narrow diameter coupled w/ direct injection of
sample & short reaction times, minimizes the extent of
lateral diffusion
2. coiling in these small tubes generate a force w/c helps cut
down diffusion
MAJOR DISTURBANCES
1. significant carry over problems
2. wasteful use of CF reagents
3. analyzer needs to be reconfigured when analyzer is to be
measured
Examples:
1. Bayer/ Technicon Instruments corp
 AA: auto-analyzer
- 1st automated analyzer by technicon
- CF: single channel segmented on the analyzer w/
approximately 40 tests/hr
Batch analysis – processed in concert as a grp/ batch is the
sane analytical analysis
SMAC: sequential non selective analysis; BAR code for ID
Sequential analysis – samples are processed sequentially rather
than in bacth
- samples enter the system one after another are processed &
results are issued in the same order
SELECTIVE/ DISCRETIONARY ANALYSIS
- samples can be processed by any indirect method/
combination of methods available in the given analyzers at
the discretion of the operator
- measurement or absorption photometry using a stationary
flow cell or filters for isolation
CENTRIFUGAL ANALYSIS
mid 1960 – Dr. Norman Anderson at Oak Ridge Natl lab
early 1970 – miniaturized version of space exploration program =
chemical analysis on outer space
spin off technology from NASA outer space research
Principle:
- centrifugal force transfers the reagent outward through
capillary channel mixes them w/ the serum sample then
transfers them into separate cuvette (LS: Xe lamp)
- measurement of A/ Fluorescence (if fluorometric
technology s being performed) as reaction proceeds in the
cuvette rotating at the periphery of the thick Teflon rotor
disc (4”)
- rotor: heart of the system in each individual reaction
chamber
- A/ F is monitored by a vertically mount photomultiplier
above the rotor; the light source is below the edge of the
rotor edge
- Electrical signals from PM – changes in the contents of
cuvette – binary digit code – Transfer & mask computer
memory
DRY SLIDE TECHNOLOGY/ THIN FILM TECHNOLOGY (closed
system)
- Eastman Kodak Ektachem – Vitros Recent J&J (1976-77)
- Primary example of dry slide technology
- great innovative approach for chem. Analysis: Kodak
- An example of a unit test concept: each slide contains
rgts/ all materials necessary for a single analyzer
- Exploits major areas of expertise layering of chemicals on
a film & rgts prep
reportable result efficiency (FPY: first pass yield)
- Reagent slide is stored in fridge/freezer temp – depending
on the test thus allows major shelf life & provides a high
degree of reproducibility
- Close system – assay parameters are set by
manufacturers & ref purchasing of reagents from the
manufacturer because of a unique container format
- Reagent can be purchased from a variety of sources or
versions (assay parameters may be modified by
purchaser)
Layering: Anderson
- spreading reagent, indicator & support (only liquid
sample)
- complete discretionary stated rate of output – 120 tests/
hr = 20uL serum
- sample size: 10uL – suitable for pediatric analysis
- Outstanding example of application of technique
(multiple thin film) developed for/ purpose
(photography) to completely differentiate ultra-micro clin
chem. Analysis
Unique aspects:
1. only fluid in the system is serum/ plasma
2. 1st : microvolumes of sample/ reagents on slide/ day
chemical analysis
3. 1st : incorporate computer technology extensively into
its design
4. drop-wise dispensing of the sample replaces the exact
measurement of the sample volume
Reflectance Spectroscopy
- replaces transmission spectrophotomoetry w/c measures
the intensity of colors by determining the proportion of a
beam of light that is reflected off the colored area
- Na & K are made in disposable ISE chip & flat Ag AgCl
electrodes
- The potential layer produced between the 2 half cells is
proportional to the concentration (activity of the
electrolyte + Na & K in the sample)
3 different analysis modes:
1. discretionary
2. single-test batching
3. STAT
 spreading layer – accepts the sample
- porous network – sample spread over a defined area
- capillary network serves as a sieve (large molecules—
proteins—are removed)
 one or more reagent layer – alters the aliquot
- contains all the chemicals necessary for the specific
reaction in a complex assay, several rgts are
successfully layered
 scavenger layer – special section
- removes materials present on the sample w/c might
interfere w/ the reaction taking place in the reagent
layer
 indicator layer – optional
- continuous dye or some othe type of indicator w/c
reacts w/ the product reactions taking place in the
reagent layer forming a colored complex
 support layer – on w/c the entire is instructed
- clear plastic w/c remains transparent when the colored
complex is being measured
RANDOM ACCESS ANALYZER
 - a system where any specimen can be analyzed in any
sequence w/ regard to the initial order of the specimen
- should analyze Px sample for only those constituents
specifically ordered (test selectivity)
- flexible enough to accommodate any reasonable
combination of requests for a single Px
- STAT samples are provided so emergency assays can be
carried out in a reasonable time by momentarily
interrupting the normal sequence of Px analysis
- System – automatically returned to normal operation
after STAT request has been filled
- Capable of incorporating new tests into the analytical
scheme by addition if the appropriate reagents & simple
reprog. parameters
- Functioning both speed & flexible req. in today’s lab
DISCRETE ANALYSIS
- whereby each specimen handled as a separate process
in its own dedicated reaction vessel (output omentalized)
- each sample has its own physical space in w/c the
individual chem. Reaction takes place, often used by
simultaneous analysis & accompanying reagents
Simultaneous analysis
- more than one analysis is performed on a sample at the
same time
- exemplify multiplicity of strategies in effect is a
multichannel analyzer
- type is capable of running multiple tests one sample at a
time or multiple samples one test at a time
- can be regarded as a logical replacement of:
1. manual pipetting
2. hard movement of tubes
3. mixing
4. incubation, etc
- most popular versatile analyzer
- ex: Dupont automated clinical analyzer (ACA)
- 1970
- 1st discrete analyzer
- Sequential analysis (Anderson)
REAGENT TEST PACK TECHNOLOGY
- heart of the system
- small plaster pouch contains all the reagents necessary
of a single analysis
- recent technology added ISE system for electrolytes
measuring Na, K, Cl, CO2
 throughput – max # of tests that can be performed by an
analyzer for a given period of time (1hr) –below 200/hr
 dwell time – min time from initial sampling to prod of result