HEMOSTASIS

Prepared by:
John Mark S. Capuyan, RMT
 HEMOSTASIS
• Maintenance of blood flow within the
vascular system
• Includes:
1. Keeps the circulating blood in its fluid state
2. Produces clot to stop the bleeding
3. Dissolves the clot as the wound heals
 STAGES OF HEMOSTASIS
I. Primary Hemostasis
A. Vasoconstriction
B. Platelet Plug Formation
II. Secondary Hemostasis
A. Stable Fibrin clot Formation
B. Inhibition of Activated Coagulation factors
 COMMON TESTS FOR
PRIMARY HEMOSTASIS
• Platelet Count
• Bleeding Time
• Clot Retraction
• Rumple-Leede Test
• Platelet Aggregometry
 PLATELET COUNT
*Reasons why platelets hard to count:
1. Platelets adhere to foreign surfaces
2. Platelets easily disintegrate
3. They are hard to differentiate from debris
4. Platelets are unevenly distributed in the blood
because they tend to clump

Normal Value (General) = 150, 000 – 450, 000 /mm^3

• Indirect Methods
- Platelets are counted in relation to 1,000 RBCs in the
blood smear
- NOT RELIABLE
Formula:
(RBC count/ 1000) x No. of Platelets = Plt count (/mm^3)
• Direct Methods
- Whole blood is diluted with platelet diluting fluid in
an RBC pipet and counted in a hemacytometer

A. Light Microscopy Methods

Rees and Ecker’s
Guy and Leake’s
B. Phase Microscopy Methods
 Rees and Ecker’s
• Diluting Fluid composed of: BCB, Na citrate,
Distilled H20
• 4 corner large squares (as in WBC count)
• Formula:
Plt ct. (mm^3) = Plt ctd. x 10 x 200
4
 Procedure
1.Rinse RBC pipet first with R & E fluid by sucking in and out the DF
2.Suck blood to 0.5 mark of RBC pipet
3.Suck diluting fluid to 101 mark
4.Shake the pipet for 3-5 minutes
5.Discard few drops
6.Charge the counting chamber and let it stand at least 3 minutes
7.Count the platelets in the 4 corner large squares
8. Use the formula
 Guy and Leake’s
• Diluting Fluid composed of: Crystal violet, Distilled
H20, Formalin (40%)
• All the 25 intermediate squares in the central
large square (as in RBC count)

• Formula:
Plt ct. (mm^3) = Plt ctd. x 10 x 200 x 1
 Procedure
1.Suck the DF to mark 1 of RBC pipet
2.Suck blood to 0.5 mark of the pipet
3.Suck diluting fluid to 101 mark (DF, blood, DF)
4.Shake the pipet for 3-5 minutes
5.Discard few drops
6.Charge the counting chamber and let it stand at least 3 minutes
7.Count the platelets in all the 25 intermediate squares in the central
large square
8. Use the formula
 Phase Microscopy
1. Brecher-Cronkite Method
DF: 1% Ammonium oxalate
Procedure: same as in R and E EXCEPT that
platelets are counted with the use of
Phase contrast microscope
2. Unopette
3. Tocantin’s
4. Nygard’s method
5. Walker and Sweeney’s method
6. Van Allen’s Method (reported in percent)
• Platelet Count Estimation
- Examine the thin area of the slide using OIO
- Normal (Wedge) blood smear should demonstrate
approximately 8 to 20 platelets per field

For the estimation of platelet count:

a. Scan 10 OIO fields for the number of platelets
b. Average number of platelets/OIF x 20,000 = EPC per uL
Estimate may be reported using the following table:
PLATELET ESTIMATE OF: REPORT AS:
0 to 49,000/ uL Marked decrease
50,000 to 99,000/ uL Moderate decrease
100,000 to 149,000/ uL Slight decrease
150,000 to 199,000/ uL Low Normal
200,000 to 400,000/ uL Normal
401,000 to 599,000/ uL Slight increase
600,000 to 800,000/ uL Moderate increase
Above 800,000/ uL Marked increase
 BLEEDING TIME
• Screening test for vWD and
disorders of Platelet
function
• Time it takes for a standard
wound to stop bleeding
• Not affected by the
coagulation mechanism
 Duke Method
• Finger is punctured (3mm deep) using a sterile
lancet
• Start timer as soon as soon as the first drop of
blood appears
• Blot drop of blood with filter paper every 30
seconds (paper must not touch the wound)
• Stop timer as soon as bleeding stops.
• N.V. = 2 to 4 mins
 Ivy Method
• Use sphygmomanometer (inflated to 40
mmHg) at patient’s upper forearm (above the
elbow)
• Cleanse an area on the volar surface of the arm
with 70% alcohol. Wipe dry with sterile gauze.
• Hold skin tightly by grasping the underside of
the arm firmly.
• Make 2 incisions (2mm deep and 2 mm long)
avoiding any subcutaneous veins.
 Ivy Method
• Start the timer and blot the drop of blood from
each puncture site on 2 separate filter paper
every 30 secs.
• Stop timer as soon as bleeding stops
• Release pressure cuff.
• Record bleeding times of the 2 puncture sites
and report the average of the 2 results
• N.V. = 1 to 7 mins.
 Template Bleeding Time
• A modification of the original Ivy Bleeding Time
test
• Used a template containing a standardized slit in
place of the disposable lancets
• Template has been replaced by several
commercially made devices (Simplate, Surgicutt)
• Gen. ref. range = 2 to 9 mins.