DOCUMENT RESUME BD 200431 0 TITLE Medical. Service Climi Laboratory ProcedureHematologi INSTITUTION Department of the Air Force, Washingon,t, Department of the'Army,- Washington, D.C. EEPORT 110 AFM-150-51; -TH-B-227-4 PUN:. DATE 5 Dec 73 NOTE- 151p.; Contains colored pictures which may reproduce Well AVAILABLE FRO Superintendent of Documents, U' S . GovernmentPriatii Office, aashington, DC 20402 (Stock No. 00-020-00525-W; $3.55). IDES PRICE MF01/Pd07 Plus Postage. DESCRIPTORS , *Allied Health OCcupations: *Armed Forces;_Biology;-- Government Publications; Health Occupations; Higher Education; Instructional Materials; naberatory Manuals: *LaboratorY Procedures; 'Medical Education; *Medical. Technologists; riaterials-zr, Science IDENTIFIERS- *Hematology .ABSTRACT Presented are laboratory studies focusing onblood- cells and theomplete scheme of blood coagulation. Forked is'the basin for the following types :of laboratory operations: (1) distinguishi4.the morphology of normal and abnormal blood cells; (2) measuring theapcentrations or number of blood cells; (3) measuring concentration a** abnormalities'detecting of heloglobin; (4) . measuring defecesin coagulation; and (5) performing a few specific disease-related-tests which involve blood cells. Types, of equipment needed, actual stkavise perormance of tests, and reagents needed,= as well as established, minimum levels of accuracy are included. lAuthor/CS) ********************************* ****** -*4***** roductions dupplied by EDRS are the.best can be from th6 original docdment. ****************************************** -- D&;ARTM-ENTS. OF THEAIR FO10E_ AF MANUAL -160-51 _ . THE 'ARM TM- 8-227-4 Washington DC 20330_ 5 December 1973- Medical Service CLINICAL LABORATORY PROCEDURES-1 The purpose of this-manual is to present the laboratoi-y steadiesw concern the blood cells and the complete: scheme of blood coagulation. It forms thebasis -for the following types- of laboratorli operations(1) clighiguishing the Morphology of normal' and abnormalblood. cells, ,(2)imeasuring:.--ti the concentrations.or number of bloodcells; (3) measuring concentra- tion driOr,.t4 tecting abnormalities of hemoglobin; -(4)measuring defects in.boagulation; and (5) pei-forining a Lei- specific disease-related testa which involve-blood cells. It enumerates types of e uipment needed, actual stepwise performanceof teSts,..and= reagents- needed, as well as e'tablishes rniiijmum levels ofaccuracy: 1 -ChaPtdr..-1Blood Section ATbe Composition of, Blood. Introduction- -- , . t 1-1 eneral Cellular Structure 6 1-2 'Cellular Constitu'ents. 1-3 Liquid Constituents _ L 1-4 1-2it ection BFormation of Blood Cells _mbryonic Heinatopoiesis, 1-5 1-2n r ostnatal Hematopoiesis_ 1-6,-, 1-3 on CNormal Cell Mtu eneral Features_ Cytoplasm ;Nucleus' SeCtiorl: DAbliormal General Features Cytoplasm Nucleus. Section' E--Functions Introduction 1-13 1-6 Erytfirocytes 1-14 1-6 Leukocytes' _ 1 1-15 _ 1-7 Platelets .= _ 1-16 1-7 Chapter 2---Material employed in He atolo`gy 'Section ALaboratory Reagents Preparation _ 2-1 2-1 Labeling Reagent Containers '2-2. 2-1 Safety Precautions- . _ _ _ _ _ .- Supersedes AFM 169751,1 May 1962 and TM 8 -227 -4, 1 Maych 1963. (For summary of -evised, deleted, or added.rhaferial; se.--siga:6,De page.) OPR: SGPSE (14repar0 by SHCS/MST)' DISTRIBUTION (Page 7-10) -AFM 60751/TM -2 7 5 Dec mbei 1973. page Section BLaboratory Glassware Diluting .Pipets 2-2 Unopettes - , 2-3 Calibration of Sahli Pipets _ 2-6. 2-3 Blood Cell Counting Chambers'.. __ _ 2-7 . 2-4 , Cleaning -_ , 2-8 2-5 7Preparation of Siliconized cilassware _ _. 2-9 2-6 Section CCompound Microscope Introduction = 2-10 , Magnification 2-11 . Illumination = 2-12- Focusing 2-13 Care-of the Microscope = 2-14 es 2-15 Section D-Centrifuges Types ; 2716 2-10 Precautions 2-17 t 2 -14 Section EAutomated Equipment Introduction 2-1a 2 -12 Dilutors-'_ - - = 2119 2-12 I 2-12 Slide Stainer =t _ Counting Devices - -,...2,21 2-12 IL Hemoglobinometer 2-22 2-17.. Fibrometer 2-23 2-17 , Hematological Systems 2-24 .217 Maintenance of Automated Hematological Equipment 9_95 2-18 Chapter 3Basic Hematological Procedures Section ACollection of-Blood Specimens' Introduction VOnipuncture Capillary- Puncture Anticoagulants Section BPreparation and Staining of-Blood Smears Introduction . Slide Meth-etl Coverglass Method Section CBone Mprrow Specimens, Introduction _ Processing of Bone Marrow Specimens for Exa inaion Chapter 4Morphology of Blood Cells Section A---General Information -Basic .Concepts Of Cell M.orphology 4-1 General Rules of Cell Identification _ 41 Section BErythroeytes IntrOduction 4-1 Erythrocytic Series 4-2 4-2 Variations in Erytocyte§ = AFt1L-1600,17I'M December --- 973 - iii pier Seetion C-Leukocytes Granulocytic Series 6 -15 Agrant31putes 4-7 .--LymphoeyticBeries _ 4-8 Monoelytic -SerieS:- 4-9 Plaamoc3-iic/Series = 10 VarrationS-of Leukocytes ion1:1-=Thrembocytes Introduction_ 4-14 Megakaryocytic Series 4-13 4:45 Chapter' 5Method§ in Hematology -Seetion -AManual Cell Counts i-1 5-1 Red Blood Cell Count (Hemacytomete 5-2 51 White-Blood-Cell Count (HemacytOmeter) 5-3 5-6 Total Eosinophil Count 5-4 /5-8 Absolute Basophil Count (Cooper a- 5 i 9: Reticulocyte _Count 5-6 0 - Cerebrospinal Fluid Cell Counts _ 5-7 -- 5--11 Semen Analysis r =8 5712 Section B---Hernatecrit Introduction- . 5-9 5-14 Madrohernatocrit (WintrObe) , 5-10- 5-14 kg Microhematocrit -- ( - . 5-11 5-16 ,- Section CErythroyte Sedimentation --Rate--. IntrOduction _ _, _: 5-12. 5i16 Detern ination of Sedimentation Bate (Wintrobe-Landsbe 5-13 5-47 Seetion D rnoglobin General nformation 5-14 -18 Coinpoun s of Hemoglobin 5-15 - 51_8 Variatio of Hemoglobin 5-16 L18 Hemoglo nometry 5-17- 5718 Cyanmeth moglobin Method 5-18_ 5-18 Detectieof Hemoglobin S andon S Sickling Hemoglobins' 5-19 5-29 Demonration of the Sickle Cell Phenomenon _ 5-20- 5-21 obin Electrophoresis (Cellulose Acetate) 5-2k 5-22 Fetal Hemoglobin .(Alkali Denaturation Test) = 5-22 5-24 Section EExamination of Blood Smears Introduction I 5-23- 5-25 Examination of Peripheral Blood Smears 5-24 b-26 Examination of Bone Marrow Smears 5-25 5-28 Section FErythrocyte Indices and Fragilitp_Tests Erythrocyte Indices . 5-26 5-28 Erythrocyte Osmotieirragility Test (Dacie ) _ 5-27 5-29 Ham' Test for Erythrocyte Fragility 5-28 5-.31 Section GDemonstration of L.E. Cells General information 5-29 5-32 Rotary Bead Method .= 5-30 5-32 v AVIV 160-51/TM S-227 1973 Section H--Special Stains .. Kaplow) ,- 5-31 - :5-L33 Peioxidase'Stain 1 LeukocyteAlkatine*Phosphatase K(Kapp')l 2 5734 Heinz Body Stain 5-33/ 545 Siderocyte Stain, __ I, 5 z34 - 5-36 Section IMisepllaneous Teas' . - ScreeningTest for Glucose-6-Phosphate Dehydrogenase Deficienty 5-35 5-36 Shipment of Cytogenetic Specimens. 5-36 5-37 Chaptor 6--Blood tCoagulation Section AIn tfOduction HemOstilsis -6=1 Coagulation 2 SectiO,BCoagulation Factors Blood. Factors Plafelet -Factors Fibrinolyfic Factors Section--CThe _Coagulation Mechanism Introduction TheModern Theory CoagulatiOrt Ihhibitors . Section pCoagulatidli Studies Introduction 6-9 6-5 Blood Collection 6710 6-5 Bleeding Tirne = 6-11 6-6 Whole- Blood 'Clotting Ti 6,-12 6-7 Clot Retraction Test 6.13 .6-8 Tourniquet Test 6-14 6 -8 Partial Thromboplastin Time (Activated 6-15 6-9 Qpe-Stage Prothrombin Time 6-16 6-10 Prothrombin Consumption Time 6-17 6-11 Thromboplastin -Generation Time 6=18 -6-12 Thrombin Time 6-19 6-14 Fibrinogen Assay (Semiquantitative) 6-20* 6=15 Plasma Recalcification Time - 6-21 6-16 Detection of a-Circulating Antidoagulant 6-22 -6-16 Detection of Factor XIII Deficiency = 6-23- 6-17. Platelet Count. (Rees-Ecker) 6-24 .6-17 Platelet Count (Phase 6-25 6-18 Macroglobulia Distilled Water Screening Test 6426 6-19 Cryoglobulin Scteening Test 6-27 .6-29 e Chapter 7Quality Control Introduction 7-1 7-1 Definition of Terms 7-2 7-1 -Statistical Evaluation of Hematology* 7-3 7-2 Control in Blood = Collection 7-4 72.6 Control in Hemoglobin Deterrnrtions 7-5 Control in cell Counting (Hemacytometer) 7-6 7-7 Control in Electronic Counting 7-7 7-8 AFM 160-51/TM. , 227 ember 1973 _ram? ., ?age Control-in -Hernatocrit Determination .7 7-8 Control in Differential Counts . _ 7L9 7-9- COntrol, in Coagulation Studies 7-10 -7-9 Control, in Other Teats : 7-11 7-9 Stun/nary, 7-12 : 7-9 Attachments 1.Bibliography A-1-1 2.Glossary A2-1 3.-Prefixes, Suffixes, and Stern only Used in. Hematology: = A3-1 4.Index A44- Figures 1-1.Typical Cell Structure 1-2 1-2.Hernatopoietic Syeterti in anAdult 18,---20 Yea of Age 1 1-3.Development of Blood Cells Hematological Pipets . 2-2. _ Unopette System 2-3.Rulings on a Hemacytorneter, 2-4.Compound Microscope.. 2-5. u. Microhernatocrit Centrifuge and Microcrritocrit Reader =- 2-1.1- 2-6.Automatic Dilutor . -2-13 2-7.Automatic Slide Stainer 2-8.Coulter Csunter Model FN 2-15 2-9,Schematic tIsiagiarn of the Coulter Counter 2-16- 2-1N. Schematic Diagram of a Photoelectric Particle Counte 2-16- 241. Fibrometer 2-18 2-12. Coulter Model S _ 2-19 -1.Site of Venipuncture _ 3-1._ 3-2.Venipuncture Procedure 3-3 Vacutainer System _ _ - 3-5 3-4.Capillary Puncture Procedure 3 -6 3-5.Slide' Method for Preparation of Blood Films 3-10 3-6.Coverglase Method for Prep_ aration of Blood Films -13 4- 1.Erythrocytic Series 4-2.Variations in Erythrocytes 4-3.Granulocytic Series _ 4-4.Lymphocytic Series and Monocytic Series 4-1 Plasmocytic Series 4=13 4-6.Variation's in Leukodytes 4-15 - 4-7.Megakaryocytic Series, _ 4-10'' 5-1.Processing Blood for Manual Cell Counts 5-2 '5-2.charging the Hernacytbrneter 5-3.Hemacytometer Counting Chamber. Ar ed1 5, are used to count red blood cells _ 5-4 5-4.Hemacytometer Counting Chamber.