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Plasma Coagulation Factors: 1 Factor I Fibrinogen 2 Factor II Prothrombin 3 Factor III Thromboplastin 4 Factor IV Ca+2 5 Factor V Proaccelerin 6 Factor VI Va 7 Factor VII Proconvertin 8 Factor VIII Antihemophilic Factor A 9 Factor IX AHF B (Christmas factor) 10 Factor X Stuart Prower 11 Factor XI Plasma thromboplastin antecedent 12 Factor XII Hageman 13 Factor XIII Fibrin stabilizing II. Intrinsic, Extrinsic and Common Coagulation Pathway Intrinsic pathway: Reaction system that begins with factor XII and culminates in fibrin polymerization Screened by: Activated partial thromboplastin time (APTT) Coagulation factors involved: XII Pre-K HMWK XI IX VIII X V Prothrombin Fibrinogen Extrinsic pathway: Tissue factor pathway Screened by: Prothrombin time test (PT) Coagulation factors involved: VII X V Prothrombin Fibrinogen Common pathway: Common path of extrinsic and intrinsic pathway Coagulation factors involved: X V Prothrombin Fibrinogen 1

III. Normal Values (CU-SI) Hemoglobin Male 14-18 gm% 140 -180 gm/L Female 12-16 gm% 120-150 gm /L 1gm Hb = 1.34 ml O2 Specific gravity of blood = 1.053 1gm Hb = 3.47 mg Iron 1% Hct = 0.34 gm% Hb = 107,000 RBC / cu mm Male 42-52 vol% 0.42-0.52 Female 37-47 vol% 0.37-0.47 Male 4.5-6.0 M / cu mm 4.5 -6.0 X 1012/L Female 4.05.5 M /cu mm 4.0-5.5 X 1012/L 5000 10,000 / cu mm 5-10 X 109 /L Segmented Neutrophil Lymphocytes Neutrophilic band (stab or staff) Monocyte Eosinophil Basophil Wintrobe: Male Female 60-70% 25-35% 2-6% 2-6% 1-4% 0.5-1% 0-10 mm/hr 0-20 mm/hr 0.60 -0.70 0.25-0.35 0.02-0.06 0.02-0.06 0.01-0.04 0.005-0.01

Hematocrit RBC WBC (x 10,000)

(x 50) Differential count


Absolute eosinophil count Reticulocyte count ( 10) Platelet count (x 2,000 Rees-ecker mtd) (x 1000 Unopette mtd) OFT (x 171) -> conversion factor

Westergren: Male 0-15 mm/hr Female 0-17 mm/hr 150-300 cells / cu mm Adults 0.5-1.5% 5-15 X 10-3 Babies 2.0-6.0% 20-60 X 10-3 200,000-400,000 cells / cu mm 200-400 X 109 /L Griffin Sanford Initial hemolysis 0.42 -0.46% Complete hemolysis 0.30-0.34% Dacies mtd Initial hemolysis Complete hemolysis Dukes Ivys (2-3mm puncture) Copley lalitch (6mm puncture) O.O Dale and laidlaws or Capillary tube 0.50-0.55% 0.35-0.40% 2-4 min 1-7 min <3 min 2-4 min

72-79 mmol/L 51-58 mmol/L 86-94 mmol/L 60-68 mmol/L

Bleeding time

Clotting time

Clotting time

Lee and white or 7-15 min Whole blood Drop or slide 2-4 min 10-14 sec 23.4-36.2 sec 8-14 sec Castor oil or 15-45 min Hirschboeck mtd Stefanini or test tube Begins within 1 hr Ends within 18-24 hrs

Prothrombin time APTT Thrombin time Clot retraction time

Capillary resistance test

Mac farlane 44-67% Tourniquet or 1+ (1-10 petechiae) Rumpel leedes or Hess test Suction or >4 petechiae at 200 mm Hg Petechiometer

IV. Tabulation of Methods Blood collection Skin puncture Venipuncture Syringe method Evacuated tube method Two slide or wedge method Two coverslips or Ehrlichs two cover glass method Spinners method Automated method Sysmex SP-100 Staining dish method Staining jar or DIP method Automated method Hemastainer automatic slide stainer Hema-Tek 1000 slide stainer Hema-Tek 2000 slide stainer Sysmex SP-100 and Beckman Coulter Colorimetric methods A. Direct method Tallquist or scale Dares hemoglobinometers Acid hematin method (Sahli hellige or Sahli adams) Alkali hematin 3

Smear preparation



B. Photoelectric or indirect matching Oxyhemoglobin Cyanmethemoglobin or hemoglobin method Carboxyhemoglobin Gasometric method A. Van slyke oxygen capacity Specific gravity A. Copper sulfate Chemical methods A. Kenneys B. Wongs Hemoglobin electrophoresis Hematocrit Macro methods A. Wintrobe B. Hadens modification C. Van allen D. Sanford magath E. Brays Micro methods A. Adams Automated counter A. Coulter counter B. Autoanalyzer ESR Macro methods A. Wintrobe and Landsbergs B. Westergren C. Graphic or cutler D. Lizenmeier E. Brays Micro methods A. Micro Landau B. Smith micro C. Crista or hellige-vollmer Automated methods A. Mini Ves B. Ves Matic C. Ves Matic 60


Absolute eosinophil count

Randolph technique Pilot eosinophil count Direct eosinophil count by Friedman Microscopic methods A. Dry method B. Wet method C. Miller disk method Automated method A. Sysmex R-1000

Reticulocyte count

Platelet count

Direct methods A. Rees-Ecker B. Nygrads C. Guy and Leakes D. Brecker Cronkite E. Van allens F. Unopette Indirect methods A. Dameshek B. Fonios C. Olefs (best indirect)


Rapid Screening test Dacies method Griffin Sanford method Quantitative test Unopette method Dukes Ivys (2-3mm puncture) Copley lalitch method (6mm puncture) O.O Capillary blood methods A. Drop or slide B. Capillary tube or Dale and Laidlaws Whole blood or Lee and White Method Qualitative A. Castor oil or Hirschboeck method Quantitative A. Stefanini or test tube B. Mac farlane Tourniquet or Rumpel-Leedes or Hess Suction or Petechiometer

Bleeding time Clotting time

Clot retraction time

Capillary resistance test

V. Diluting Fluids RBC Dacies fluid or Formol citrate (best rbc diluting fluid) Hayems Gowers Toissons Bethells NSS 3.8% Sodium citrate 1-3% Acetic acid with gentian violet 1% HCl Tuerks Phloxine Pilot Randolphhs


Absolute eosinophil count