HEMA REVIEWER!!!!!
I. Capillary Fragility Test
Principle:
Determination or measure of capillary
integrity to not allow blood to escape
(Hemorrhage).
Procedure:
1. Apply Sphygmomanometer
(100mmhg)
2. Maintain for 5 minutes
3. Release pressure
4. Observe after 2 minutes for
Petechiae (small-Reddish spots that
are <3mm)
Results:
Females: (+) if more than 10
Petechiae is observed
Males: (+) if 5 Petechiae is observed
*Presence of Petechiae not necessarily
abnormal.
II. Bleeding Time (Ivy Method)
Principle:
Measurement of the time it takes for a
standard wound to stop bleeding.
Procedure:
1. Apply Sphygmomanometer
(40mmhg)
2. Choose the puncture. Find a volar
surface within the arm which is free
of blood vessels usually the most
muscular part is suitable.
3. Disinfect area of the puncture
4. Puncture 3 equidistant holes
5. Start the Timer when the blood
begins to appear.
6. Wait for 2 minutes
7. Begin blotting. Using Filter paper
at the sides of the wound.
8. Continue blotting with a 30 second
interval
9. Stop blotting when Blood no longer
appears in the filter paper.
10. Report Results
Reference Value: 2-8 Minutes 5. Use WBC squares in HPO. Follow
the inverted L rule (Right and up not
*If exceed 15 minutes Stop procedure
counted)
and report as >15 Minutes.

IV. Platelet Count: Direct Method

by Rees and Ecker
Principle:
Measurement of Platelet count by
staining platelet and counting via
Hemocytometry

Procedure:
1. Dilution
-RBC pipette (0.5-1-101)
Reese and Ecker fluid up to .5 mark Computation:
*Components Cell Count X DF X VCF
-Sodium Citrate- Prevent coagulation =/ Cumm
- Formalin- Fixes platelet and prevent DF= Vol of Fluid in bulb
premature hemolysis Vol of Sample used
= (101-1)
-Brilliant Cresyl Blue- Allows platelet
.5
to be observed =200

VCF= Desired Volume (1)

2. Fill Pipette with sample up to 1 # Of Squares (4) X Vol of
mark square (0.1)
3. Dilution Fluid again up to 101 =2.5
Mark
Reference Value:
4. Charge the Hemocytometer make 150,000 – 450,000 cumm
sure not to overfill or underfill each 150-450X109/L
chamber
V. Indirect Method of Platelet Count VII. Coagulation Time (Lee & White
(Fonio’s Method) Whole Blood Clotting) “Tube Method”

Principle: Principle:
Derivation of Platelet counting relative to Time it takes for Blood to clot after contact
RBC count with a negatively charged surface

Procedure:
1. Skin puncture
-Disinfect area
2. Drop 14% Mgso4 in finger
3. Puncture within drop
4. Allow Mixture
5. Smear with Wright’s Stain
6. Collect Blood in Capillary tube for
determination of HCT and divide by 3 and
divide by 3 to get RBC count
7. Count 10 fields in OIO
PLT COUNT=
# of PLT counted in 10 Fields
Approx Value in 10 (1000) X RBC count
=X109/L
Procedure:
1. 3 tubes of 13X100
2. Label as “3” “2” “1”
3. Sample Collection via Venipuncture
4. Time as soon as blood appears in the hub
of the syringe.
5. Collect 5ml of blood
6. Remove Needle
7. Dispense Sample Blood must touch the
glass (Activates XII) 1 ml Per tube
following Tube 3, Tube 2 and Tube 1.
8. Water Bath 37C for 5 minutes
9. After 5 minutes observe tube 1 by tilting
in a 45 angle (Clot formation).
10. If tube 1 has clotted observe tube 2 then
3 after 30 seconds interval
*Don’t tilt tube 2 if tube 1 has not yet
clotted.
Results:
Linearity- 1 minute
Reference Value: 5-15 Minutes

VIII. Slide Method

VI. Platelet Estimate Procedure:
1. Make sure glass is clean
Principle: 2. Perform Finger puncture
Count Platelets in 10 OIF multiplied to 3. 3 Dome Shape
Platelet Estimation Factor 4. Start checking for fibrin thread after 2
minutes and check every after 30 seconds.
-PBS is Used (EDTA)
Normal Values: 2-8 Minutes
Plt Count=
Plt in 10 X19
10
= 109/L
IX. Prothrombin Time XI. Clot Retraction Test (Macfarlane
Method)
Principle: Measures the Extrinsic
coagulation factors along with the common Principle: Measures the amount of time it
Pathway takes for the blood clot to pull away from
the walls of the test tube. Evaluates Blood
Procedure: Platelet Disorders
1. 100Ul of plasma added 100Ul Tissue
thromboplastin Procedure:
1. Collect a 6ml of sample
2. Cuvette with Steel ball added with 10Ul 2. In a graduated centrifuge, dispense 5ml
plasma while allowing the blood to touch the glass.
3. Add an applicator stick
3. Incubate 4. Incubate 2 hours
5. Remove stick with clot
4. 100Ul reagent 6. Centrifuge 3000 rpm for 3 minutes
7. Compute serum.
5. Reading well
Computation:
Normal Value: 10.8- 13.8
Volume of serum-Packed cell volume
5ml
X. Activated Partial Thromboplastin X100%
Time
CRT=%
Principle: Measures the Intrinsic
coagulation factors along with the common Normal Value= 43-65%
Pathway
Results
Procedure: 1. Characteristic of the clot
1. Steel Ball with cuvette is added with 50Ul - Solid Conical Clot
sample - Amorphous Clot

2. 50Ul of PTR/ RGT is added 2. Degree of Retraction

- Complete
3. Incubate - Partial

4. Reading well and add 50Ul Cacl2 3. % of Retraction

Normal Value: 25-48 Seconds

XII. Lupus Erythematosus Preparation

Principle:
Determination of the presence of LE cells

Procedure:
1. Blood Clot – Macerate to destroy Red
cells

2. Fill wintrobe tube all to 0 mark

3. Centrifuge for 30 minutes

4. Harvest Buffy coat and add serum then

Mix.

5. Smear dry Stain with Wright’s Stain