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Clinical Hematology Laboratory

Hematology is the branch of medicine, that is concerned with the study of blood, blood forming organs and blood diseases. It includes study of etiology, diagnosis, treatment, prognosis and prevention of blood diseases .

After the completion of this presentation we will know about:
What is hematology and its purpose.
hematology laboratory.
Blood and its compositions and collections
Hematology lab equipment's
Some hematological tests , disease and hazards too.

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Clinical Hematology Laboratory

  1. 1. Prepared & presented By: Rojal Neupane Ruru Rikham Magar Shivangi Verma Yamuna Nepal Tracy Khadka Presented To: Tapeshwar Yadav (Lecturer) BMLT, DNHE, M.Sc. Medical Biochemistry Department of Laboratory Medicine, Nobel College of Medical Sciences, Kathmandu, Nepal CLINICAL HEMATOLOGY LABORATORY
  2. 2. After the completion of this presentation we will know about:  What is hematology and its purpose.  hematology laboratory.  Blood and its compositions and collections  Hematology lab equipment's  Some hematological tests , disease and hazards too . 2
  3. 3. In Greek, the word hematology is derived from hem(e)= blood logy = study of Father of hematology : William Hewson Hematology is the branch of medicine, that is concerned with the study of blood, blood forming organs and blood diseases. It includes study of etiology, diagnosis, treatment, prognosis and prevention of blood diseases . 3
  4. 4. It encompasses the study of blood component and coagulation . It includes : 1. Analysis of concentration, structure, and function of cells and their precursors in the bone marrow. 2. Analysis of chemical constituents of plasma or serum intimately, linked with blood cell structure and function. 3. Studies of functions of platelets and proteins involve in blood coagulation. Changes in one or more of the characteristics mentioned above may produce or cause hematological diseases or manifestations. 4
  5. 5.  Hematologist plays an important role to find out the causes of blood borne diseases by providing the physician the required laboratory results.  Hematologist helps the patients to get better treatment by providing the accurate test results to the physician.  It deals with routine determination of total number of cells in circulation, hemoglobin concentration and differential counts of leukocytes based on study of the stained blood smear.  Stained blood smear helps in detecting morphological abnormalities of various cell in seen in peripheral blood circulation. 5
  6. 6.  Blood volume: 5-6 liters (8% of total body weight)  Slightly alkaline, PH: 7.53-7.45  Specific gravity : 1.052-1.060  Viscosity : 4.5 times more viscous than water  Temperature : 36-38˚ C  Osmotic pressure: average 25 mm of Hg  Color: red due to presence of hemoglobin in RBC  Taste: salty 6
  7. 7.  Transport of oxygen from lungs to tissue and carbon dioxide from tissue to lungs.  Transport of metabolites, nutrition and metabolic wastes.  Acts as a buffer and regulates acid-base balance.  Regulation of body temperature.  Regulation of water balance.  Maintains ion balance.  Defense against infection by WBC and antibodies. 7
  8. 8. Blood is a specialized connective tissue which circulates in a closed system of blood vessels. Blood is composed of blood cells suspended in a pale yellow colored plasma. A: Cellular substance - 45% Erythrocytes or Red Blood Cells (RBC) Size: 7 – 8 micrometer Life span: 120 days Function: Transport of oxygen and carbondioxide 8
  9. 9. 9
  10. 10. Leukocytes or White Blood Cells (WBC)  Granulocytes Neutrophil Size: 12-15 micrometer Life span: 4-8 hrs Functions: phagocytosis and destruction of bacteria Eosinophil Size: 12-16 micrometer Life span: 4-8 hours Function: Kills parasites larvae, regulate mast cells, response to inflammation 10
  11. 11. Basophil Size: 8-10 micrometer Life span: 4-8 hrs Function: mediate inflammatory response  Agranulocytes Lymphocytes Size: 6-8 micrometer Lifespan: day to year Function: Key cell in immune system Monocytes Size: 16-22 micrometer Life span: 10-20 hours Function : Precursors to macrophages histocytes 11
  12. 12. 12
  13. 13. Thrombocytes or platelets Size: 2-4 micrometer Life span: 7 -10 days Function: Blood clotting 13
  14. 14. B: Liquid intercellular substances (plasma) - 55% Plasma contains  Liquid i.e. water (90-92%)  Plasma protein 7% and includes Albumin, Globulin, Fibrinogen, Prothrombin  Inorganic Salt 0.9% includes sodium Chloride, Sodium bicarbonate salts.  Traces of organic materials like urea, uric acid, creatinine, cholesterol, etc.  Trace of nutrient material (from digested food): Monosaccharaides (mainly glucose), Amino acids, fatty acids, glycerol, vitamins  Trace amount of hormone and enzymes .  Plasma carries dissolved respiratory gases like oxygen, carbondioxide, nitrogen and also antigens and antibodies. 14
  15. 15. 15 Fig.: Components of blood.
  16. 16. 16
  17. 17. Collection for a small number of routine tests blood may be collected by finger or ear lobe prick how ever for performing large number of routine test a larger quantity of blood is collected from vein puncture. There are mainly two ways for collection of blood. Collection of Capillary Blood by skin puncture This method can be use to perform tests for which only few drops of blood is needed. 17
  18. 18. Site for collecting capillary blood:  Finger tips ( ring or index finger)  Heels  Ear lobes 18
  19. 19. Methods  Wipe and clean the site with cotton swab containing 70% alcohol or spirit  Prick with the sterile lancet. The cut should be deep enough (3 mm) so that blood flows freely without squeezing the punctured site.  Discard first drop of blood and wipe with dry cotton and then let it to form round drop freely.  Take Blood for required test and press the punctured site with cotton swab to stop further bleeding. 19
  20. 20. Tests performed by using capillary blood  Hemoglobin  Platelets count  Bleeding Time, Clotting Time  Blood grouping and Rh typing  Thick smears for malarial parasites  Blood count  Reticulocyte count 20 Note: Capillary blood should be used only when it is not possible to obtain blood from veinpuncture. Capillary blood may give erroneous results.
  21. 21. Collection of blood by Venipuncture About 2-3 ml of blood is collected in either EDTA or double oxalate bulb. Prothrombin time is determined from blood collected in trisodium citrate. Site of obtaining venous blood Anti-cubital vein is the best site for collection. However radial vein, dorsal vein, ankle vein can also be used. 21
  22. 22. 22
  23. 23. Methods  To locate the vein the arm of patient is kept warm and tourniquet is tied to the upper arm.  Ask the patient to make fist.  Skin over the vein is cleaned with cotton swab soaked in 70% alcohol ( ethanol) and let it dry.  Inspect needle and syringe for blockage.  The patient’s arm is gripped tightly and thumb of other hand is used to grip the skin taut.  The vein is penetrated with the angle at 30-40˚  Insert the needle in prominent vein by fixing the vein and withdraw the appropriate amount of blood by pulling the piston slowly. 23
  24. 24.  When blood is collected in syringe, then release the tourniquet and remove needle from vein  A cotton ball is held firmly over the venipuncture site as soon as needle is removed.  Transfer the collected blood into labeled anticoagulated vials by removing the needle (which prevents hemolysis) and mix well 24 Note: Air must not be pumped into the vein.
  25. 25. 25 Fig.: Venipuncture
  26. 26. Tests performed by blood collected by veinpuncture:  Hemoglobin  Erythrocyte count  Leukocyte count  Packed cell volume  Reticulocyte count  Platelets count  ESR by Wintrobe’s method 26
  27. 27. Colour Anticoagulants Uses Red or yellow - For serum Lavender EDTA ( Na2 or k2) Whole blood for CBC Blue Sodium citrate ( Liquid) Whole blood for ESR and coagulation tests Green Heparin Plasma or whole blood Grey Sodium Fluoride Plasma for Blood glucose 27
  28. 28. 28 Fig.: Tubes
  29. 29. 29
  30. 30. Microscope It is used to see microorganisms that Cannot be seen by our naked eyes. • Micro-hematocrit centrifuge it is used to determine the specific product value of blood cell and separate micro-blood from micro-solution . 30
  31. 31.  Water bath Made from container filled with water. Used in lab to incubate samples in water maintained at constant temperature. Differential cell counters It is used in DLC cell count. 31
  32. 32.  These are used to perform complete blood counts, erythrocyte sedimentation rates (ESRs), or coagulation tests. 32
  33. 33.  Slide-staining racks It helps in staining many slides at a time, as when we put many slides on a rack we can pour dye simultaneously on all the slides . Stop watch 33
  34. 34.  Micropipettes A micropipette is used to transfer small volumes of liquids in chemical, biological and medical laboratories. Pressing on a plunger button at the top of the micropipette will pull the liquid in, and a second press will dispense it. 34
  35. 35. 35 It is used to measure a blood sample and to determining the red cell count
  36. 36.  White blood cell (WBC) pipette It is used for WBC count .  Cell counting chambers It is used to measure the concentration of RBCs , WBCs and also bacteria , virus and other pathogen in blood. 36
  37. 37. 37  used for ESR (Wintrobe's method), PCV, haematocrit, etc. Westergren’s tubes used for ESR (Westergren's method)
  38. 38.  Similarly some other glasswares are:  Pasture pipettes,  Glass slides, 38
  39. 39.  Disposable syringe, needles and lancets  Tube stands 39
  40. 40.  Drabkin’s reagents  RBC diluting fluid  Staining solution  Wright’s stain  Leishman’s stain  Platelet diluting stain  3.8 gm/dl trisodium citrate  4.0 gm/dl EDTA solution  Brilliant cresyl blue solution 40
  41. 41.  Routine hematological test: Hemoglobin  Method: Cyanmethemoglobin method  Normal values Male: 13- 18 gm/dl Female : 12-16 gm/dl children upto 1yr: 11-13 gm/dl chlid (10-12) yrs : 11.5-14.5 gm/dl  Increase values seen in hemoconcentration due to loss of body fluid, heart diseases, polycythemia.  Decreased value is observed in anemia 41
  42. 42. Total Leukocyte Count (TC)  Normal values: 4000 to 11000 /mm3  Increase in WBC count is known as leukocytosis.  Decrease in WBC count is known as leukopenia. Differential Leukocyte Count (DC) Neutrophil: 40 – 75%  Increased condition (Neutrophilia) - observed in leukocytosis  Decreased condition (Neutropenia) - observed in bacterial infections and other conditions such as anemia, suppression of bone marrow by various drugs and radiation. 42
  43. 43. Lymphocytes: 20-40%  Increased condition (Lymphocytosis)- observed in infection such as mumps, measles, influenza and other chronic infections  Decreased condition (Lymphopenia)- observed in acute stages of infection and excessive irradiation Eosinophils: 1-6%  Increased condition eosinophilia – observed in asthama, parasitic infestation, in chronic inflammatory diseases. Monocytes: 2-10%  Increased condition monocytosis – observed in tuberculosis, malaria, typhoid and kala-azar 43
  44. 44. Basophils: 0-1%  Increased condition basophilia – observed in chronic myeloid leukemia RBC count (erythrocytes)  Normal value: 4.5 - 5.5 millions /mm3  Increased value observed in hemconcentration due to burns, dehydration, etc.  Decreased value observed in polycythemia, old age, pregnancy, etc. 44
  45. 45. Platelets count  Normal value: 250,000 – 500,000 / mm3  Increased condition (Thrombocytosis) – observed in polycythemia, chronic myelogenous leukemia  Decreased condition (Thrombocytopenia) – observed in prolonged bleeding, aplastic anemia, acute leukemia, immune thrombocytopenia Reticulocytes count  Normal range: Adults 0.2 – 2% Infants 2-6%  Increased count indicates increase activity of bone marrow (Hemolytic anemia or acute blood loss)  Absence or low count indicates bone marrow suppression (Aplastic anemia) 45
  46. 46. 46
  47. 47. PCV (Packed Cell Volume)  Method : Microhematocrit method  Normal values: Male 40-52 % Female 36-48 %  Decreased values observed in anemia, hydremia (excessive fluid in the blood as occurs in pregnancy)  Increased values observed in Polycythemia, Dehydration, Congenital heart disease. RBC indices  MCV (Mean Cell Volume) = PCV x 10/RBC in millions Normal Range 82-92 fl Increased value seen in Macrocytic anemia Decreased value seen in Microcytic anemia 47
  48. 48.  MCH (Mean Cell Hemoglobin) = Hb X 10 / RBC in millions Normal Range 27 – 32 pg Increased value seen in Macrocytic anemia Decreased value seen in Hypochromia  MCHC (Mean Corpuscular Hemoglobin Concentration) = Hb x 100 / PCV Normal range 32-36 % Increased value seen in Spherocytosis Decreased value seen in Hypochromic anemia 48
  49. 49. Study of morphology of blood cells • In different anemia and other diseases, the morphology and adequacy of blood cells in blood smear may show certain significant changes. ESR (erythrocytes sedimentation rate)  Method: Westergren’s method  Normal value: Male 0-15 mm after an hour Female 0-20 mm after an hour  Increased in all conditions where there is tissue breakdown or entry of foreign protein in blood. The changes are however not diagnostic of any specific diseases. 49
  50. 50.  Coagulation tests: Bleeding time(BT)  Method: Duke’s method  Normal range: 1-5 minutes  Prolonged bleeding time is generally associated with thrombocytopenia. Bleeding time helps to detect vascular defect and platelet disorder. Clotting time (CT)  Method: Capillary or Lee –white method  Normal range: 4-9 minutes  This method is generally useful in severe clotting disorder. 50
  51. 51. Prothrombin time(PT)  Method: Quick’s method  Normal range: 14 ±2 secs.  This measures the quality of the extrinsic path way of coagulation. Activated partial thromboplastin time (APTT)  Method: Quick’s method  Normal range: 35-40 sec  APTT measures the efficiency of intrinsic and common coagulation pathway. This test is performed to diagnosis hemophilia that involves the deficiency of the clotting factors. 51
  52. 52. Thrombin time (TT)  Method: Quick’s method  Normal range: 15- 20 secs  Increased value is observed in decrease of fibrinogen concentration, presence of dysfunctional fibrinogen or high concentration of fibrin - fibrinogen degradation products or presence of heparin. 52
  53. 53.  Special tests:  Detection of fetal hemoglobin Marked increase in level of HbF are observed in thalassemia, sickle cell anemia and in other congenital disease and acquired hematological conditions.  Detection of blood parasites Diseases like Malaria, Leishmania (Kala-azar), Filaria, Trypanosoma, etc. can be diagnosed by observing blood parasites and their stages under microscope.  Microscopic examination of bone marrow In different type of anemias and diseases such as leukemia, multiple myeloma microscopic examination of bone marrow is done. 53
  54. 54. 54
  55. 55.  Anemia Types of anemia  Iron deficiency anemia  Hemolytic anemia  Aplastic anemia  Megaloblastic anemia  Sideroblastic anemia  Sickle cell anemia 55
  56. 56.  Thalassemia  Abnormal hemoglobin and hemoglobinopathies  Leukemias  Tumor of lymphoid tissue  Parasitic infections of blood  Malaria  Filaria  Kala-azar  Trypanosoma 56
  57. 57.  Transmission of blood borne diseases and other infections.  Inhalation of harmful reagents and chemical causing serious illness.  Corrosive chemicals causing serious injuries and burns.  During the collection of sample one may prick oneself.  Combustible chemical may catch fire during tests.  Broken glasses may cause cuts, bleeding and infections while handling damaged slides or coverglass or pipetting with broken ends. 57
  58. 58.  Regular use of Personal protective equipments (PPE) like aprons, gloves, glasses etc..  Use of safety signs for harmful and corrosive chemicals.  Separate designated areas for collection and storage of samples.  Safe pipetting (no mouth pipetting) and dispensing.  Safe use of syringe and needles.  Use one hand while needle recapping.  Necessary course of vaccination must be taken. 58
  59. 59. 59 Fig.: Use of PPE Fig.: Safety signs
  60. 60.  Disposing of samples in designated areas.  Proper disposal of syringes and needles.  Use of needle destroyer for destroying needles.  Place reusable glassware, plasticware, specimen container etc in 5% of hypochlorite solution, autoclave, wash and reuse.  Store radioactive waste for 3 months before decontamination and disposal as medical waste. 60
  61. 61. 61 Fig.: Waste management
  62. 62. 62
  63. 63.  Anemia : condition below normal value in PCV , RBC count  Anticoagulant : Chemical used to inhibit clotting of whole blood , the liquid portion of the sample harvested plasma  Basophil : a class of granulocytic leukocyte that promotes the inflammatory response  Erythrocytes : RBC counting granules  Hematology : the science that deals with the morphology og blood and blood forming tissue and with their physiology and pathology 63
  64. 64.  Heme : a non protein iron containing portion of hemoglobin  PVC : packet cell volume or hematocrit.  Plasma : fluid protein of the blood in which cells are suspended. 64
  65. 65.  TEXT BOOK OF MEDICAL LABORATORY TECHNOLOGY (Third edition) by P.B. Godkar and D.P. Godkar  TEXT BOOK OF MEDICAL LABORATORY TECHNOLOGY (First edition Volume II) by R.K. Gupta and B.K. Yadav 65
  66. 66. 66