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Coagulation - Lab

This document describes a laboratory exercise on blood physiology. The exercise involves several experiments: [1] testing physical and chemical factors that influence blood coagulation in vitro, [2] determining coagulation time in humans using capillary and slide methods, and [3] performing blood typing. The objectives are to discuss blood typing principles, factors affecting coagulation, proper blood handling, and relating results to clinical conditions.

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Guia De Guzman
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155 views7 pages

Coagulation - Lab

This document describes a laboratory exercise on blood physiology. The exercise involves several experiments: [1] testing physical and chemical factors that influence blood coagulation in vitro, [2] determining coagulation time in humans using capillary and slide methods, and [3] performing blood typing. The objectives are to discuss blood typing principles, factors affecting coagulation, proper blood handling, and relating results to clinical conditions.

Uploaded by

Guia De Guzman
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Laboratory Exercise #6

Blood Physiology

Blood is the main fluid of the body that maintains homeostasis. It distributes oxygen and
nutrients to the different tissues, plays an important role in immunity, acid –base balance and
hemostasis.

Objectives:
1. Discuss the principles in blood typing and cross matching.
2. Discuss factors that will favor and will not favor blood coagulation.
3. Demonstrate proper handling of blood and fluids in extraction.
4. Correlate results with common clinical conditions affecting blood typing, coagulation and
bleeding time.

I. Blood Coagulation (in Vitro)


Coagulation is the change in the physical state of the blood from liquid to solid. This
experiment would show the student some factors influencing coagulation and the technique of
determining coagulation in man.
A. Some Physical Factors Influencing Blood Coagulation
Each of the following test tubes is assigned to a student.
1. Test tube #1 – containing nothing. Serves as control for physical factors.
2. Test tube #2 – paraffinized and contains 2 ml of paraffin oil
3. Test tube #3 – contains a wire for continuous beating of blood once placed inside
4. Test tube #4 – contains nothing but will be placed in a beaker of ice
5. Test tube #5 – contains nothing but is placed in a beaker with water at a To of 43oC
6. Test tube #6 – contains a piece of gauze
B. Some Chemical Factors Influencing Blood Coagulation
1. Test tube #7 – contains nothing and serves as control for chemical factors
2. Test tube #8 – contains 0.5ml of CaCl2
3. Test tube #9 – contains 0.5ml of 10% sodium citrate
4. Test tube #10 – contains 0.5ml of sodium oxalate
5. Test tube #11 – contains 0.5ml of 10% sodium fluoride
6. Test tube #12 – contains 0.5ml of 25% magnesium sulfate
7. Test tube #13 – contains 0.5ml of heparin
8. Test tube #14 – contains 0.5ml of minced muscle tissue

Proper blood extraction will be done by the students on volunteers. Blood will be
distributed according to the number of test tubes. Students are encouraged to line up based on the
above mentioned order for proper documentation. Each students should have their own timer to
record the exact time while observing for the outcome of the experiment. Timer is started when
the blood gets into each test tube.

The blood in the test tube should not be disturbed (except for Test tube #3), but
every 15 seconds, the tubes should be slowly tilted to determine whether coagulation has already
set or not. The end point for coagulation can be taken as the time when the tube can be inverted
without displacement of the blood. In some test tube, it may be necessary to observe as long as
30 minutes. Tabulate your results.

Tube No. Blood let at (Time) Blood coagulated at Coagulation Time


(Time)
1

10

11

12

13

14

Questions:
1. What is the mechanism involved in each of the above experiment to explain the effects on
blood coagulation?

2. What are the common anticoagulants in vitro? In vivo?


II. Blood Coagulation in Man
The purpose of this experiment is for the student to learn two methods of determining
coagulation time in man: the capillary and the slide methods. Both methods will be performed on
each student.

A. Capillary Method
After properly sterilizing the subject’s fingertip, prick it with a sterile lancet/
pricker to secure a free flow of blood (Do not squeeze the finger). Note the time of the
first appearance of blood. Discard the first drop. Let the next drop of the blood seep into
the capillary tubing (capillet) by touching the drop of blood to the capillary tube held
almost parallel to the finger, which is in horizontal position. At intervals of 15 seconds,
break off a short segment of the capillet and gradually draw the pieces apart. Look for
fibrin threads between the two broken ends. This fibrin indicates the onset of coagulation.
Note the time of the initial appearance of the fibrin thread.

B. Slide Method
Place a good-size drop of blood on a slide by pricking the subject’s fingertip after
proper sterilization. Note the time when the blood first appears on the finger. At intervals
of 15 seconds, draw a pin from the center of the blood drop towards the periphery and
observe if any fibrin strands go with the pin. Note the time of initial appearance of this
fibrin strand. The time when the blood has clotted minus the time when the blood is shed
equals the coagulation time.

Questions:
1. Why is the determination of coagulation time in man necessary?

2. Why is it not advisable to squeeze the finger after pricking?

3. Which of the two methods give a longer coagulation time? Why?


4. What is the normal coagulation time in man when the blood is extracted from the skin? When
extracted from the vein? Why is there a difference between the two? Explain.

5. Differentiate between coagulation and bleeding time.

6. Tabulate your results below.

Subject Coagulation Time


Capillary Method Slide Method

Mean (Average)
III. Blood Typing
One day before the performance of the experiment, the student should report to the
laboratory for blood withdrawal or extraction. One drop of his blood will be mixed with 1.0 ml
of NSS to provide the red cell suspension, while the rest will be allowed to clot in order to obtain
the serum.

Questions:
1. What is clot retraction? Give the mechanism involved.

2. What is the physiologic significance of clot retraction?

Technique of Blood Typing


1. Get two clean slides, one for the standard serum and the other for the standard cells.
Label the slides.
2. Place the drop of standard serum A on the left half of the slide and one drop of the
standard serum B on the right half. To each of these, add one drop of your cell
suspension. Observe macroscopically at first then study microscopically. Note if
agglutination has occurred.
3. On the others slide, place one drop of standard cell A on the left half of the slide and one
drop of standard cell B on the right half. To each of these, add one drop of your serum.
Observe for agglutination.

4. Write your observation as follows:


Serum A + Cell X = ____________ Possible Blood Types: ________ and _________
Serum B + Cell X = ____________ Possible Blood Types: ________ and _________
Therefore, the agglutinogen/s present in my red blood cell is/ are: ________________

Cell A + Serum X = ____________ Possible Blood Types: ________ and _________


Cell B + Serum X = ____________ Possible Blood Types: ________ and _________
Therefore, the agglutinin/s present in my red blood cell is/ are: __________________

I therefore conclude that my blood type is: ____________


Questions:
1. How do you detect agglutination macroscopically?

2. What will you do to differentiate rouleaux formation from true agglutination as seen under the
microscope?

3. In the ABO grouping, what agglutinins can agglutinate the following agglutinogens: A? B? Is
there an “O” agglutinogen?

4. What are some of the uses of blood typing?


5. Does human blood contain agglutinogen and agglutinin other than those shown in this
experiment? Enumerate them, if any.

6. If your blood type is “AB,” could any of your offspring have a blood type of “O?” Explain.

7. What is the purpose of cross-matching blood?

8. Discuss positive major and minor reactions in cross matching.

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