Documente Academic
Documente Profesional
Documente Cultură
By:
Ahmed Makboul Ahmed
Assistant Lecturer of Clinical Pathology, SECI, Assiut University
Outlines:
- Platelet structure.
- Functions of Platelets.
- Overview of normal platelet function.
- Platelet function tests:
o Overview.
o Light transmission platelet aggregometry
(LTA).
o Other platelet function tests.
INTRODUCTION:
Stages of Hemostasis:
1. Primary Hemostasis:
- Platelets and the injured vessels interact to form the primary hemostatic plug,
which is a clump of platelets.
- It only temporarily stops the bleeding and is very fragile.
2. Secondary Hemostasis:
- The fibrin-platelet plug is then formed by a series of interactions involving the
coagulation factors (coagulation cascade).
- The fibrin stabilizes the clot covering the hole in the vessel.
3. Fibrinolysis:
- The process of removing the clot once the wound has been healed.
PLATELET STRUCTURE:
Normal human platelets are:
Size: Small in size (0.5 x 3.0 μm)
Shape:
- Discoid in shape have a mean volume of 7–11 fL.
- Anucleated.
Number: They circulate in relatively high numbers (between 150
and 400 x 109 /L).
Life span: Their lifespan is approximately 10 days (9 – 12 days).
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2
Platelet Ultrastructure
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1. Peripheral Zone:
Function:
- Adhesion and aggregation.
- Responsible for cell's negative charge.
a). Microtubules:
- Located beneath the cell membrane of resting
platelet.
- Maintain discoid shape.
b). Microfilaments:
- Mediate contractile events.
- Growth modulators:
PDGF.
TGF-β.
Platelet factor 4 (PF4).
- Coagulation factors:
Factor V.
High-molecular-weight kininogen (HMWK).
C1 inhibitor.
Fibrinogen.
Factor XI.
Protein S.
Plasminogen activator inhibitor-1 (PAI-1).
1.2. Transport:
- After collection, samples should be transported to laboratory at room temperature.
- Tubes should not be subjected to vibration, shaking, vortexing, continuous mixing,
or agitation.
- Samples should be tested between 30 min and no more than 4 hr from blood
collection.
2. PROCESSING:
2.1. Citrated PRP:
- Prepared by centrifugation at 800 – 1000
rpm for 15–20 minutes.
- Platelet Count of PRP:
• Platelet count of PRP is performed and
should be 200-400 x 10⁹/L.
• If platelet count is > 400 x 10⁹/L,
concentration can be adjusted with
autologous PPP.
- PRP is added to a cuvette at 37C and
preincubated for up to 5 minutes.
- The PRP is stirred at a recommended speed
(e.g., 1000–1200 rpm using a magnetic stir
bar) to allow platelets to come in contact
with each other.
2.2. Instrument calibration:
Aggregometers are calibrated for light transmission using
autologous PPP and PRP:
- vWF antigen and activity studies and factor VIII activity assay can be performed if
vWD is suspected.
In vWD, there is increased response to low-dose ristocetin. This occurs in:
Increased low-dose RIPA with patient plasma mixed with normal platelets is
consistent with type 2B vWD
2. Genetic studies: can also be used to distinguish type 2B vWD from platelet-
type vWD.
b). Abnormal aggregation response only to collagen agonist may be
noted if there is:
o defect in collagen receptors (GPIa-IIa or GPVI).
6. Plateletworks®:
Plateletworks® is a standardized platelet counting ratio technique,
based upon comparing platelet counts within a control EDTA tube
and after aggregation with platelet agonists within citrated tubes,
could theoretically also be applied to the diagnosis of platelet
disorders, although experience with this assay is limited.