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EXPERIMENTS ON

BLOOD

SABANDO/SABANDOSR/SABANGAN/SADANG/SALARZ
ON/SALAS


BLOOD
COAGULATION
Tube No. Blood let at Blood coagulated Coagulation time
at
1 2:09 PM 2:14:07 5minutes 7secs
2 2:09 PM 2:17 8mins
3 2:09 PM 2:17:42 8mins 42secs
4 2:09 PM 2:15 6mins
5 2:09 PM 2:14:54 5mins 53secs
6 2:09 PM 2:09:40 40secs
7 2:09 PM 2:14 5mins
8 2:09 PM -
9 2:09 PM -
10 2:09 PM -
11 2:09 PM -
12 2:09 PM -
13 2:09 PM -
14 - Theoretical -
15 - Theoretical -
Test Tube 2 - paraffin oil causes delay in blood
clotting, it is said that blood clotting is dependent
on vascular surface; if it is not water repellent
(non-hydrophobic) is suitable for blood
preservation in fluid condition. In general the
vascular lining is believed to be composed of
acidic polysaccharides materials.

Test Tube 3 defibrination, Removal of fibrin from


the blood, usually by means of constant agitation
while the blood is collected in a container.
Test Tube 4 - chilling causes changes in the
platelets' outer membrane. When the
temperature drops, some of the molecules in the
membrane, such as cholesterol and another
molecule called sphingomyelin, clump into
distinct islands in the membrane called "lipid
rafts." Some proteins attached to the surface,
including some that carry signals from the cell
surface to the inside, are also collected into these
rafts.
Test Tube 5 - causing irreversible reduction of
coagulation activity in the plasma. At the same
time thromboplastin- and PTT-reagent are
imparied in their coagulation acitvity by an
increase in temperature.

Test Tube 6 gauze


The active ingredient found in QuikClot is a
naturally occurring inert mineral called kaolin,
which has been known for decades to clot blood.
Kaolin does not contain animal or human proteins
or botanicals.
Test Tube 8 - The addition CaCl2 of solution into
the blood led to partial disruption of red cell
aggregation and decrease of hematocrit in the
blood.

Test Tube 9 - removes calcium ions (Ca 2+) from


blood plasma. It also prevents blood from clotting.
Note that by removing calcium ions from the
blood, sodium oxalate can impair brain function,
and depositcalcium oxalatein the kidneys.
Test Tube 10 like CaCl2, sodium citrate also
removes calcium ions (Ca2+) from blood plasma. It
also prevents blood from clotting.

Test Tube 11 Sodium Flouride, the red cells are


prevented from using any more glucose. In other
tubes, the red cells are still functioning and they
continue metabolizing the glucose which can
make theglucose levelsappear lower than they
really are in the body. (glycolytic)
Test Tube 12 Thrombin time was prolonged by
addition of MgSO4, and fibrin concentrations

Test Tube 13 Heparin, Highly


sulfatedglycosaminoglycan, is widely used as an
injectableanticoagulant, and has the highest
negativecharge densityof any knownbiological
molecule. Heparin binds to the enzyme
inhibitorantithrombinIII (AT)
Test Tube 14 Dicumarol, Is a naturally
occurringanticoagulantthat functions as a
functionalvitamin Kdepleter.

Test Tube 15 - From inactive actin to activated


actin causing viscosity of the blood hastens
coagulation.
GUIDE
1.
QUESTIONS:
What is the mechanism involved in each of the above experiment to
explain the effects on blood coagulation?

Tube #2 Delayed coagulation

Tube #3 Defibrination

Tube #4 Delayed coagulation


Tube #5 Fastened coagulation

Tube #8 Anti-coagulant
Tube #9 Anti-coagulant

Tube #10 Anti-coagulant


Tube #11 Anti-coagulant

Tube #12 Act as fibrinogen

Tube #13 Prevents formation of prothrombin


Tube #14 Anti-coagulant

Tube #15 Hastens coagulation


BLOOD
COAGULATION
2. What are the common anti-coagulants in vitro? In vivo?

vitro vivo
Oxalate Heparin
Citrate Dicumarol
Na fluoride Anti-thrombin
Heparin
Magnesium sulfate
Calcium chloride
Coagulation Time
(Clotting Time)
Clotting Time is the time
required for blood to form a clot
in vitro.
The basis for the test is that
whole blood will form a solid
clot when exposed to a foreign
surface such as a glass tube.
It is a rough measure of all
intrinsic clotting factors in the
absence of tissue factors.
Clotting Time - Slide
Method

This test is sensitive


to the factors
involved in the
intrinsic pathway
The expected range
for clotting time is 4-
10 min.
Clotting Time - Capillary method

Normal value is 3 to
10 minutes.
Determination of coagulation time in man is necessary

To prevent excessive blood loss


To check for low levels of blood clotting
factors
To check for low levels of Vitamin K
Normal Coagulation Time

Skin
4-6 mins
Trauma is induced
Vein
10-15 mins
No trauma, no tissue factor
RESULTS
SLIDE METHOD CAPILLARY METHOD

45 SECS CLOTTING
TIME 45 SECS CLOTTING
TIME
BLOOD TYPING
A drop of Serum A was placed
on the left half of the side and
Serum B on the right side.

1 drop of of cell suspension was


placed on the slides and was
observed.
RESULTS

AB

A
BLOOD
TYPING
1. What is clot retraction?
After the clot has been formed, the activated
platelets incorporated in the clot rearrange and
contract their intracellular actin/myosin
cytoskeleton.
The intracellular actin network is connected to the
internal part of the integrin IIb3 fibrinogen
receptor.
Following coagulation, the external part of IIb3
will have bound to the fibrin network of the clot, and
as a result of platelet contractile force on the fibrin
network, the formed clot will compact on itself and
hence reduce its total volume.
BLOOD CLOT
Dissolution
of Clots
When damage is
repaired, activated
factor XII causes
activation of
kallikrein

o Kallikrein
converts
plasminogen to
plasmin
Plasmin
digests fibrin,
dissolving clot
Clot retraction & Repair

As clot is compacted, fibroblasts


(stimulated by platelet-derived growth
factor -PDGF) rebuild the wall while
endothelial cells (stimulated by vascular
endothelial growth factor -VEGF) multiply to
restore the lining
Clot retraction
& Repair
Clot retraction occurs w/in 20-60
minutes
Platelets contain actin & myosin,
thrombasthenin
o Platelets pull on surrounding fibrin
strand & squeezes serum out of
the mass
o Serum: plasma minus clotting
proteins
BLOOD TYPING

What is the physiological significance


of clot retraction?

The physiological function is to clear


the obstructed vessel for renewed
blood flow.

AGGLUTINATION
AGGLUTINATION: The clumping together of
red blood cells or any particulate matter
resulting from interaction of antibody and its
corresponding antigen.

ANTIBODY: A protein substance secreted by


plasma cells that is developed in response to
and interacting specifically with an antigen.

ANTIGEN: A substance recognized by the body as


being foreign,which can cause immune response.
Forward and reverse
grouping normally yield
strong(positive
4)reactions.
Agglutinins and Agglutinogen
Agglutination versus
Rouleaux
Causes of Rouleaux
Serum containing altered protein
proportions, abnormal proteins, or high
concentrations of fibrinogen.
Patients who have received plasma
expanders of high molecular weight,
intravenously injected contrast materials,
or certain drugs may also have serum that
induces rouleaux formation.
Cell typing may also be affected if the cells
are suspended in their own serum.
HOW TO
DETECT TRUE
AGGLUTINATIO
N?
SALINE TEST
PROCEDURE
Set up the serum confirmation test as described
under ABO GROUPING - SERUM CONFIRMATION.
Centrifuge but do not resuspend the cells.
Using a dispo pipet, draw off and discard the
supernatant serum.
Add two drops of physiologic saline to each
tube.
Gently resuspend and observe macroscopically
for agglutination using the lighted view lamp.
Grade and record results.
INTERPRETATION

Removing the serum and replacing with


saline dilutes the interfering proteins so
that rouleaux formation is prevented, but
does not alter agglutination due to Anti-A or
Anti-B. Rouleaux should disperse in the
saline; true agglutination will remain
agglutinated.
Interpretation
Negative Positive
saline test saline test
Uses of Blood Typing

Blood Transfusion
Paternity Determination
Medical
Importance

Prenatal Care
Forensic
Investigation
Rh Antigen
There are six common types of Rh antigens, each
of which is called an Rh factor. These types are
designated C,D, E, c, d, and e.
A person who has a C antigen does not have the c
antigen, but the person missing the C antigen
always has the c antigen. The same is true for the
D-d and E-e antigens.
The type D antigen is widely
prevalent in the population and
considerably more antigenic than the
other Rh antigens.
Anyone who has this type of antigen
is said to be Rh positive, whereas a
person who does not have type D
antigen is said to be Rh negative.
However, it must be noted that even
in Rh-negative people, some of the
other Rh antigens can still cause
transfusion reactions, although the
reactions are usually much milder.
If your blood type is
AB, could any of your
offspring have blood
type of O?
BLOOD CROSS
MATCHING
What is blood cross
matching?
Also known as compatibility testing,
pretransfusion testing or type and
crossmatch (Type and Cross; T & C).
the final step in pretransfusion testing

a series of procedures use to give an indication


of blood group compatibility between the
donor and the recipient and to detect irregular
antibodies in the recipient's serum.

The main purpose for performing a


crossmatch is to promote (not ensure) the
safe transfusion of blood
Cross-matching falls
into two categories:
Major Cross-match :

Recipient serum is tested against donor


packed cells to determine if the
recipient has preformed antibodies
against any antigens on the donor's
cells. This is the required cross-match
prior
Minor to release of a unit of packed
Cross-match:
cells.
Recipient red cells are tested against
donor serum to detect donor antibodies
directed against a patient's antigens.
This is no longer required. It is
assumed that the small amount of
donor serum and antibodies left in a
How is cross-matching done?
To perform a cross match, a small
amount of the recipient's serum is mixed
with a small amount of the donor RBCs.
The mixture is then examined under a
microscope. If the proposed transfusion
is incompatible, the donor RBCs are
agglutinated by antibodies in the
recipient's serum.
What is the importance of doing
cross-matching?

The purpose of the crossmatch is


a final check of ABO compatibility
and, to a lesser extent, detection of
unidentified antibodies.
What is the purpose of doing cross
matching blood?

To ensure the compatibility of the


donor and recipients blood. To be
able to prevent severe transfusion
reaction.
Give the significance of a positive major
reaction.

A POSITIVE major reaction indicates


the incompatibility between the
donors cell and the recipients
serum. Which indicates that the
blood should not be transfused
otherwise sever transfusion
reaction will occur and may
Give the significance of positive
minor reaction with a negative major
reaction.
A POSITIVE minor reaction with a
NEGATIVE major reaction can be transfused
slowly or incase of emergency. By
transfusing slowly, plasma of donor will be
diluted so no major reaction can occur.

What should be the results of the


cross-matching if the two blood
samples are compatible?

There will be no possible major and


minor reaction. There will no
agglutination from either test.
THANK
YOU!