The document provides a comprehensive overview of hematology, detailing its definition, purpose, and the significance of hematological tests and blood analysis. It covers topics such as blood composition, collection methods, laboratory equipment, and various hematological diseases and their tests. Additionally, safety measures and laboratory protocols are highlighted to ensure protection against potential hazards in the clinical hematology setting.

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. 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. 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. 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.  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.  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.  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. 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
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9. 9

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
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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
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12. 12

13. Thrombocytes or platelets
Size: 2-4 micrometer
Life span: 7 -10 days
Function: Blood clotting
13

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
Fig.: Components of
blood.

16. 16

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. Site for collecting capillary blood:
 Finger tips ( ring or index finger)
 Heels
 Ear lobes
18

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. 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. 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

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.  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
Fig.: Venipuncture

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. 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
Fig.: Tubes

29. 29

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.  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.  These are used to perform complete blood counts,
erythrocyte sedimentation rates (ESRs), or coagulation
tests.
32

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.  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
It is used to measure a blood sample and to
determining the red cell count

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
 used for ESR (Wintrobe's method), PCV,
haematocrit, etc.
Westergren’s
tubes
used for ESR (Westergren's method)

38.  Similarly some other glasswares are:
 Pasture pipettes,
 Glass slides,
38

39.  Disposable syringe, needles and lancets
 Tube stands
39

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.  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. 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. 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. 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. 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

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.  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. 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.  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. 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. 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.  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.
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54. 54

55.  Anemia
Types of anemia
 Iron deficiency anemia
 Hemolytic anemia
 Aplastic anemia
 Megaloblastic anemia
 Sideroblastic anemia
 Sickle cell anemia
55

56.  Thalassemia
 Abnormal hemoglobin and hemoglobinopathies
 Leukemias
 Tumor of lymphoid tissue
 Parasitic infections of blood
 Malaria
 Filaria
 Kala-azar
 Trypanosoma
56

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.  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
Fig.: Use of PPE
Fig.: Safety signs

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
Fig.: Waste management

62. 62

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.  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.  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

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