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CLOTTING TIME Slide, Wright’s, and Lee & White Method

EXPERIMENT 2
To achieve this unit a learner must:
OUTCOMES

Be able to perform different methods of blood clotting time determination.

UNIT

Be able to correlate the results to different diseases or disorders.

TEACHING AND LEARNING ACTIVITIES

Laboratory Experimentation

Clotting time is the interval between the moment when bleeding starts and the moment
when the fibrin clot thread is first seen. Bleeding time and clotting time are not the same.
Bleeding time depends on the integrity of platelets and vessel walls, whereas clotting time
depends on the availability of coagulation factors. When the blood vessel ruptures, in a few
minutes blood losses its fluidity and set into a semisolid mass called clot. This process in call
blood coagulation. In vivo, blood clots outside the body on cuts and injuries. In vivo, blood clots
inside the blood vessels. In coagulation disorders like hemophilia, clotting time is prolonged but
bleeding time remains normal.

PRE-ANALYTICAL PHASE

Blood Clotting
(Extrinsic pathway)
 Glossary of Terms

Coagulation: The process of stopping the blood flow from the wound. This process involves
the harmonious relationship of the blood-clotting factors, the blood vessels, and the fibrin-
forming and fibrin-lying system.
Hemophilia: A genetic disorder where longer clotting time due to absence of some clotting
factors.
In vivo: Process that occurs within the living organism.
In vitro: An artificial environment outside the living organism

ANALYTICAL PHASE

Materials

Patient preparation, setting up of working area and extraction of blood via capillary method
and syringe method.

A. Slide Method or Drop Method

1. Blood lancet
2. Glass slide
3. Timer / Stopwatch
4. Cotton
5. 70% ethyl alcohol

B. Wright’s Method or Glass Capillary Method

1. Capillary tube, non-heparinized
2. Blood lancet
3. Timer / Stopwatch
4. Cotton
5. 70% ethyl alcohol

C. Lee and White Method

1. Glass test tubes, 3 pieces (13 x 100mm)
2. Syringe, G21/G23 needle
3. Water bath 37° C
4. Timer / Stopwatch
 Procedure

A. Slide Method

1. Disinfect site of puncture with 70% ethyl alcohol. Air dry.

2. Puncture to a depth of 3mm using a blood lancet.
3. Start timer as soon as the first drop of blood appears.
4. Transfer the three drops of blood onto a clean glass slide. Careful not to touch the
skin.
5. Pass the tip of the lancet through the first drop of blood every 30 seconds and not
for the formation of fibrin strands. Repeat with second drop and then the third drop.
6. Stop the timer immediately as soon as fibrin strands are seen clinging at the tip of
the lance on the third drop.

Reference Range: 2-4 minutes

Procedure

B. Wright’s Method or Glass Capillary Method

1.Apply 70% alcohol to the clean finger with cotton swab. Allow it to dry naturally.
2.Prick the finger with usual aseptic precautions. Immediately start the stopwatch.
3.Dip one end of the capillary into blood drop gently without pressure.
4.Allow to fill the capillary with blood by lowering the end of fitted capillary. (Do not
stuck the blood around ¾ of its length undipped.
5. After about two minutes start snapping off small lengths of the tube, at intervals of
15 seconds, each time noting whether the fibrin thread is formed between the
snapped ends.
6. Repeat breaking at regular time intervals, till fibrin thread appears at the broken end
of capillary tube. Do not pull away the catted pieces.
7. Record the time interval between pricking finger and first appearance of fibrin
thread at the broken ends of capillary tube. That is clotting time of blood.
Diagram:
 Reference Range: 4-9 minutes

C. Modified Lee and White Clotting time

Principle of the Test: The coagulation of blood is the length of time required for a measured
amount of blood to clot under certain conditions. The test is based on the fact that when
venous blood is put into a glass tube (foreign surface), it will form a solid clot. The time
required for this response is a measure of the overall intrinsic and common pathways of
coagulation.

1. Label three, 13 x 10mm test tubes with Px’s name, and number them.
2. Perform aseptically a venipuncture using a 20-gauge needle syringe and withdraw 4
mL of blood.
3. After obtaining the blood, remove the needle from the syringe, and carefully place 1
mL of the blood in test tube #3, then 1 mL in tube #2, and 1 mL to tube #1. The last 1
mL must be discarded (follow universal safety procedures on the disposal of needles,
etc.). Start the stopwatch as soon as the blood is placed in tube #3.
4. Place the three tubes in a 37°C water bath.
5. At exactly 5 minutes, tilt test tube #1 gently to a 45-degree angle. Repeat this
procedure every 30 seconds until the test tube can be completely inverted without
spilling the content (that is, until the blood is completely clotted).
6. Record the time it took the blood in test tube #1 to clot.
7. Thirty seconds after the blood in test tube #1 had clotted, proceed with tube #2, and
repeat the preceding procedure tilting the tube every 30 seconds, until it is
completely clotted. Record the result. Repeat this procedure for test tube #3.
8. Since agitation and handling, speed up coagulation, the coagulation time of test tube
#3 is the result to be reported.
Reference Range: 7-15 minutes
 POST-ANALYTICAL PHASE
Critical Thinking

1. Enumerate conditions wherein the clotting time would be decreased.

2. Enumerate condition wherein the clotting time would be increased.

3. Cite the possible causes of falsely decreased clotting time.

4. Cite the possible cause of falsely increased clotting time.