HEMA 2 LEC: Laboratory Evaluation Primary Hemostasis
Platelets VWF AND GP IB
PRIMARY HEMOSTASIS: - __ play a key role in the rapid response to blood vessel injury by: Adhering to the endothelial wall at the site of injury - RESPOSIBLE BY __
- 1-10 uM - P2Y1 - P2Y12
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: ADP Typical Final Concentration: Platelet Membrane Receptors:
- 500 uM - TPa - TPb
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: Arachidonic acid Typical Final Concentration: Platelet Membrane Receptors:
5 mg/mL - GP Ia/IIa - GP VI
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: Collagen Typical Final Concentration: Platelet Membrane Receptors:
- 2-10 mg/mL - a2-adrenergic receptor
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: Epinephrine Typical Final Concentration: Platelet Membrane Receptors:
1 mg/mL - GP Ib/IX/V in association with von Willebrand factor
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: Ristocetin Typical Final Concentration: Platelet Membrane Receptors:
1 unit/mL - PAR1 - PAR4; GP Iba - GP V
Platelet Aggregometry Agonists, Reaction Concentrations, and Platelet Receptors: Agonist: Thrombin Typical Final Concentration: Platelet Membrane Receptors:
ALPHA2-ADRENERGIC RECEPTOR
Platelet Aggregometry Agonists: EPINEPHRINE: - BINDS WITH PLATELETS - BINDS WITH __ = IDENTICAL WITH MUSCLE RECEPTORS
EPINEPHRINE
Platelet Aggregometry Agonists: __: BINDS WITH PLATELETS; BINDS WITH ALPHA2-ADRENERGIC RECEPTOR = IDENTICAL WITH MUSCLE RECEPTORS
THROMBIN
Platelet Aggregometry Agonists: __: INDUCES THE ACTIVATION
ADP
Platelet Aggregometry Agonists: __: MOST COMMON RELEASED AGONIST
ARCHIDONIC ACID
Platelet Aggregometry Agonists: __: SUBSTRATE FOR EICOSANOID PATHWAY
40% FORMALIN CRYSTAL VIOLET
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method (Guy and Leake) - __ PRESERVES THE PLATELETS - __ AS A DYE TO IDENTIFY
- sodium oxalate - 40% formalin - crystal violet.
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method (Guy and Leake) - diluent is made of
𝑃𝑙𝑎𝑡𝑒𝑙𝑒𝑡𝑠 𝑐𝑜𝑢𝑛𝑡𝑒𝑑 𝑋 10 𝑋 200 /4
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method (Rees and Ecker) - Calculation: Platelets/uL=
- sodium oxalate - brilliant cresyl blue - formalin - distilled water
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method (Rees and Ecker) - Diluent is made up of
𝑝𝑙𝑎𝑡𝑒𝑙𝑒𝑡𝑠 𝑋 𝑅𝐵𝐶 𝑐𝑜𝑢𝑛𝑡 /1000
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) - Formula: Platelets/uL=
- Brilliant Cresyl Blue - Sodium Citrate - sucrose - formalin; - counter stain with Wright's Stain
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) - Needed
WRIGHT STAIN
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) __: FOR COUNTER STAINING
FORMALIN
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) __: TO PRESERVE THE PLATELETS
SODIUM CITRATE
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) __: TO PREVENT COAGULATION ; SUCROSE
BRILLIANT CRESYL BLUE
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Dameshek Method) __: USES __ AS A DYE
14 % MgSO4 + 1 drop of blood + Wright Stain
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Indirect method (Fonio's Method)
1. Guy and Leake 2. Rees and Ecker 3. Brencher-Cronkite Method 4. Electronic Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Under Direct Method
Aspirin tolerance Test 2
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination - Assesses the effect of a standard dose of aspirin on the Duke's Bleeding Time __ TABLETS OF ASPIRIN (10MG) = BLEEDING TIME VIA DUKES METHOD
Modified Ivy's Method 3-6 minutes
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination - Best method to assess platelets. It is considered to be the best screening test available for platelet's role in hemostasis. NV:
Duke's Method 1-3 minutes
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination - CAN BE PERFORMED IN EAR LOBE OR FINGER NV:
- Coply Lalitch Method - Adelson Crosby Method
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination - MUCH MORE TOXIC METHODS BECAUSE THE FINGER WILL BE PRICKED AND IMMERSED IN A BEAKER WITH STERILE NSS
- Coply Lalitch Method - Adelson Crosby Method 170-340 seconds
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination - involve immersion of the wound finger in a sterile NSS warmed at 37̊C until bleeding stops NV:
Macfarlane Method 170-340 seconds
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination -Same principle with Adelson-Crosby method but it only uses ear lobe as the site of puncture. NV:
3-6 minutes
TEST FOR PRIMARY HEMOSTASIS: Methods of Bleeding Time Determination -Template Method NV:
1. Borchgrevink Method 2. Salzman Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function - Under Tests for adhesion of platelets
drug-induced platelet dysfunction (aspirin, clopidogrel, antibiotics) - STOPPING FROM INTAKE IS THE BEST ACTION - uremia - myeloproliferative disorders
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Platelet Aggregation Tests - Acquired platelet function disorders
Defects of adhesion - Bernard Soulier syndrome Defects of aggregation - Glanzmann thrombasthenia Defects of secretion - Alpha or Dense Granule Deficiency - Aspirin-Like Defects
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Platelet Aggregation Tests - Hereditary platelet function disorders includes:
optical density
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Platelet Aggregation Tests - measures the changes in __ of a platelet suspension under conditions of constant temperature and continuous agitation
citrated plasma (PRP)
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Platelet Aggregation Tests - studied in suspensions of
Aggregation
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Platelet __ Tests - evaluation of suspected hereditary and acquired disorders of platelet function
2𝑁𝐷 𝑇𝑈𝐵𝐸 − 3𝑅𝐷 𝑇𝑈𝐵𝐸/ 1𝑆𝑇 𝑇𝑈𝐵𝐸 𝑋 100
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - FORMULA FOR SALZMAN METHOD
Borchgrevink Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - MUCH TOXIC TEST. ONCE THE FINGER IS PRICKED SAMPLE WILL COLLECTED IMMEDIATELY
CONTROL PLATELET COUNT PLATELET COUNT HOWEVER LOWER COUNT
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - USES 3 TEST TUBES: 1ST TUBE = 2ND TUBE = 3RD TUBE =
Salzman Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - Venous blood is aspirated directly from the vein through a bead column and into a vacutainer.
Borchgrevink Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - expressed as a percentage of the venous platelet count
Borchgrevink Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - platelets are enumerated in the capillary blood oozing from a bleeding time puncture
Salzman Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Function Tests for adhesion of platelets - test for the retention of platelets within glass bead columns
- Hirschboeck - MacFarlane - Stefanini-Dameshek
Test for Clot Retraction time
- Borchgrevink Method - Salzman
Test for Platelet Adhesiveness
Platelets
PRIMARY HEMOSTASIS: - Releasing potent anticoagulant compounds
platelet counted X 5 X 10 X 100
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method (Guy and Leake) - Calculation: Platelets/uL=
- Ristocetin - ADP - Collagen - Epinephrine
Agonist used in Platelet Aggregation Test
Afibrinogemia
Defective Clots - blood does not clot
Fibrinogenopenia
Defective Clots - clot is normal, increase red cell fall out
- Thrombocythemia (PLATELETS NOT FUNCTIONAL) Polycythemia (PANMYELOSIS)
Defective Clots - defective retraction; clot flabby/fragile -Increased red cell
Hyperprotenemia
Defective Clots - layered clot -clot may not retract or may retract poorly - rapid sedimementation of red cell
A. thrombocytopenia b. Thrombasthenia
Defective Clots -clot nonretractile or retracts poorly -clot edematous/friable
- epistaxis- NOSE BLEEDING - ecchymosis - genitourinary bleeding - gingival bleeding
Defects in primary hemostasis: - are generally associated with mucocutaneous bleeding, characterized by :
1. Duke's Method 2. Modified Ivy's Method 3. Coply Lalitch Method 4. Adelson-Crosby Method 5. Macfarlane's method 6. Aspirin tolerance Test 7. Template Method
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS - Methods of Bleeding Time Determination
a) elasticity of the cut tissue b) ability of the blood vessel to constrict and retract c) mechanical and chemical action of platelets in the formation of the hemostatic plug
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS Factors that affects bleeding time:
Bleeding Time
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS - ASSESSES THE PLATELET AND BLOOD VESSEL WALL
QUALITATIVE QUANTITATIVE
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS - INVOLVES THE QUANTITATIVE AND QUALITATIVE MEASUREMENT: __: FOCUSES ON THE FUNCTION __: FOCUSES ON THROMBOCYTOPENIA AND THROMBOCYTOSIS
Bleeding Time
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS - measures platelet plug formation
Bleeding Time
LABORATORY METHODS: TEST FOR PRIMARY HEMOSTASIS - provides assessment of platelet count and function
platelet plug (temporary)
PRIMARY HEMOSTASIS: - Serves to immediately limit bleeding through the formation of a __
Platelets
PRIMARY HEMOSTASIS: - Aggregating to form a plug
Platelets
PRIMARY HEMOSTASIS: - Providing a phospholipid surface for activated coagulation enzyme complexes
Clot Retraction Time
TEST FOR PRIMARY HEMOSTASIS: - Depends on normal number of contractile platelets, the presence of calcium and ATP, and a normal concentration of fibrinogen.
- LOW PLATELET = DOES NOT RETRACT - LOW CONCENTRATION OF FIBRINOGEN - PARA PROTEINEMIA = PROTEINS ARE COATED WITH PLATELETS
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time - ABNORMAL CRT YIELDS TO:
platelets calcium and ATP fibrinogen
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time - Depends on normal number of contractile __, the presence of __, and a normal concentration of _.
THROMBOSTHENIN
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time - INVOLVES __ = A CONTRACTILE PROTEIN FROM PLATELETS
1. Hirshboek Method or Castor Oil Method 2. Stefanini Method 3. Macfarlane Method
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods:
Hirshboek Method or Castor Oil Method PROTRUSION OR MAPLE LIKE
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - DROP OF BLOOD IN THE TUBE MUST SHOW ____ PROTRUSION = PLATELETS RETRACTED
Macfarlane Method
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - IT WILL BE CENTRIFUGE AND MEASURE HOW MUCH SERUM
SERUM = VOLUME OF SERUM X VOLUME OF WBC X 100 44-67%
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - MACRFARLANE FORMULA NV:
Hirshboek Method or Castor Oil Method
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - MEASURES THE PRESENCE OF DIMPLING PHENOMENON
Macfarlane Method 1 HR
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - USES A WIRE IN THE TUBE AND MEASURES WITHIN ___HR
15-45
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods Hirshboek Method or Castor Oil Method - RESULTS ARE TAKE WITHIN __ MINS
Hirshboek Method or Castor Oil Method 5-10 mL 20
TEST FOR PRIMARY HEMOSTASIS: Clot Retraction Time Methods - USES TUBE THAT HAS ___ ML OF CASTOR OIL AND __ UL OF BLOOD DROPPED IN THE TUBE
1. Thrombocytopenic purpura 2. Acute leukemia 3. Aplastic anemia 4. Scurvy (collagen and plt injury) 5. Toxins 6. destructive disease of the liver 7. HDN 8. Severe anemia
TEST FOR PRIMARY HEMOSTASIS: PROLONGED BLEEDING TIME
Indirect Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count - PERFORMING SMEAR
Direct Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count - Platelets are counted in a hemocytometer as in erythrocytes and leukocytes
Indirect Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count - Platelets are counted in their relationship to red cells on a fixed-stained smear.
Indirect Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count - This method is NOT RELIABLE because the results depend upon the distribution of platelets and on the red cell count.
1. Fonio's Method 2. Dameshek Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count - Under indirect method
Brencher-Cronkite Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - BEST DILUTING FLUID = AMMONIUM OXALATE
Brencher-Cronkite Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - BEST METHOD
Rees and Ecker
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - COUNTED IN THE 25 RBC SQUARES
Rees and Ecker
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - LOW COUNT
Electronic Method - Voltage-pulse counting - Electro-optical Counting
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - Red cells must first be removed from whole blood, either by sedimentation or by controlled centrifugation - Counting by
Brencher-Cronkite Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - USES 1% AMMONIUM OXALATE AS THE DILUTING FLUID
Brencher-Cronkite Method
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - USES PHASE CONTRAST MICROSCOPY
Rees and Ecker 1:20
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - USES WBC PIPETTE AND PERFORM __ DILUTION
RBC
TEST FOR PRIMARY HEMOSTASIS: Platelet Count Direct Method - USES ___THOMA PIPETTE HEMOCYTOMETER AND DILUTING REAGENT