Dr'R'Thamilselvi, MD',

1

• Dr.R.Thamilselvi, MD.,
• Associate Professor,
• VMKVMC

2
Haematology

• Study of the formed cellular blood elements
• Blood is composed of a Liquid called Plasma of
  cellular elements (RBCS, WBCS Platelets)

3
Blood plasma is the liquid component of blood, in
which the blood cells are suspended.
4
Blood serum is blood plasma without fibrinogen
or the other clotting factors.
5
How to approach

• Medical history Physical examination
• Select appropriate tests
• Cost- effective
• Specific test

6
To diagnose various disorders

• Disorder of blood cells
• RBC Eg anemia, polycythemia
• WBC Eg leukemia,leukopenia
• Platelets Eg thrombocytopenia, thrombocytosis
• Disorder of clotting factors
• Infections
• Malaria
• Babesiosis
• Microfilaria

7
Specimen collection

• Phlebotomy
• Anticoagulants
• EDTA (Lavender)-CBC
• Sodium citrate- (light blue) - Coagulation
  Profile
• Heparin (Green) Osmotic fragility

8
Collection of Blood
9
Ethylene diamine Tetra-acetic Acid (EDTA )

• This is the anticoagulant of choice for the FBC.
• di-Na and di-K salts are solids and tri-Na is the
  liquid form.
• Action - EDTA chelates calcium ions to form a
  soluble complex
• Uses
• Haematology Profile
• EDTA blood smears
• Concentration1.5mg/mL /- 0.35mg/mL
• Disadvantages
• EDTA cannot be used for coagulation studies (as
  it chelates calcium)
• Platelets can sometimes be seen to satellite
  neutrophils

10
Sodium Citrate

• This is a liquid anticoagulant.Mode of
  ActionRemoves calcium ions by forming a soluble
  calcium citrate complex.Uses
• Coagulation studies   91 - bloodcitrate
• ESR   41 - bloodcitrate
• Reticulocyte count
• Heinz body detection
• Acts as both a diluent and an anticoagulant
• Concentration0.109mg/mL
• Disadvantages
• Cannot use for FBC because of the dilution factor

11
Heparin

• This anticoagulant is an acid mucopolysaccharide
  
• Action- Prevents the formation of thrombin from
  prothrombin therefore stopping formation of
  fibrin from fibrinogen
• Uses
• Enzyme studies
• Cell cultures
• Osmotic fragility testing
• Concentration15IU/mL /- 2.5IU/mLDisadvantages
• Relatively Expensive
• Can give a blue background to blood films
• Platelet aggregation

12
Bedside Lab Tests

• Hb
• CBC
• Total WBC Count
• Total RBC Count
• Differential Count
• Platelet count
• Hematocrit
• Red cell Indices
• ESR
• Reticulocyte count
• Peripheral smear Examination

13
In cases of bleeding disorder

• Screening tests
• 1. Platelet count
• 2.Bleeding Time
• 3. Clotting Time
• Special tests
• 1. Prothrombin Time (PT)
• 2. Activated Partial Thromboplatsin Time (APTT)
• 3. Thrombin Time (TT)
• 3. Factor assay

14
Haemoglobin estimation

• Physical Based on Specific gravity
• Chemical Based on Iron content of Hb
• Gasometric-Based on O2 combining capacity of Hb
• Calorimetric-Based on colour
• Photoelectric Cynmethhemoglobin method
• Automated methods

15
Sahlis Haemoglobinometer
16
Procedure

• Take N/10 HCL in the tube up to lowest mark.
• Fill the Hb pipette with 0.02ml of blood wipe
  off the excess blood at nozzle and transfer it to
  the tube.
• Mix the acid and blood by shaking the tube well
  with stirrer
• Allow it to stand for 10 minutes to allow brown
  colour to develop (acid haematin)
• Now dilute the solution with distilled water
  adding few drops.
• Mix the solution well and match the colour with
  the comparator.
• The lower meniscus is noted and reading on the
  tube is read as gm/dl.

17
Cyanmethaemoglobin methodPRINCIPLE

• Blood is diluted in a solution of potassium
  ferricyanide and potassium cyanide. The
  ferricyanide oxidizes haemoglobin to
  methaemoglobin.
• Potassium cyanide provides cyanide ions to form
  Cyanmethaemoglobin.
• The absorbance of the solution is then measured
  in a spectrophotometer at a wavelength of 540 nm
  or in a colorimeter using a yellow-green filter.

18
Procedure

• Take 20 microlitres of well mixed anticoagulated
  venous blood in a glass tube and to this add 5ml
  of Drabkins solution. Stopper the tube, invert
  it several times to mix well. Allow it to stand
  at room temperature for 3-10minutes
• Absorbance is measured against reagent blank at
  540nm either in a spectrophotometer or in a
  photoelectric colorimeter with suitable filter.
• Hicn standard after bringing it to room
  temperature is taken and the absorbance measured
  using the same instrument.

19
Photoelectric colorimeter withgreen filter or at
a wavelength of 540nm

• Cynmethaemoglobin method

20
RESULT

• Calculated by using the formulae
• Hb gm/dl
• ABSORBANCE OF TEST / ABSORBANCE OF STANDARD X
  CONC. OF STANDARD.
• ADVANTAGES
• All forms of Hb except SHb are readily converted
  to HiCN.
• Direct comparison with HiCN standard possible.
• Easy to perform the test.
• Reagents are readily available.
• The standard is stable.

21
DISADVANTAGES

• Increased absorbance not due to hemoglobin may be
  turbidity due to abnormal plasma proteins,
  hyperlipaemia, high WBC count or fat.
• Potassium cyanide in the solutions is poisonous,
  though it is present only in a low concentration
  hence the reagent is not handled carefully.
• Explosion can occur if undiluted reagents are
  poured in the sink. Hydrogen cyanide is released
  by acidification and the gas if it accumulates
  can result in explosion.

22
Total WBC count

• Visual haemocytometric method
• Electronic method
• Instruments WBCpipette, Neubauerchamber,Lancet,
  cotton, Micro-scope, Cover slip and WBC diluting
  fluid (Turkes fluid)

23
Total WBC count

• Composition - Turks fluid
• Glacial acetic acid - 3ml to lyse RBCs
• 1Gentian violet - 2ml to stain the WBC
  nuclei
• Water - 100ml

24
Procedure- Total WBC count

• Suck the blood from finger prick or
  anticoagulated blood up to 0.5 in WBC pipette
• Wipe tip and outside the pipette
• Draw diluting fluid up to mark 11 in the WBC
  pipette
• Mix well by rotating the pipette for 2-3 minutes
• Discard 2 drops of the mixture from the WBC
  pipette
• Charge the Neubauer chamber

25
Procedure-
26
Neubauer Chamber - Total WBC count -Counting
areas (Blue)
27
Calculation-

• No. of WBCs in 4 large squares N
• Dilution 1 in 20 or 1/20 (11-1/0.5)
• Depth 0.1mm or 1/10
• No.of WBC's in 1 cu.mm Nx20x10
• 4
• Nx200
  / 4
  
• Nx50
• NORMAL VALUES
• Adults ? 4,000 to 11,000 cells/cu.mm
• At birth ? 10,000 to 26,000 cells/cu.mm
• Childhood ? 6,000 to 15,000 cells/cu.mm

28
Causes Aplastic anaemia Typhoid Drugs R
adiation therapy Cytotoxic therapy Subleukemic
leukemia
Causes Exercise Leukemoid reaction
Bacterial
and viral infection Infectious mononucleosis
29
Total RBC count

• Instruments
• RBC pipette, Neubauer chamber, Microscope,
  cotton, lancet, spirit, RBC diluting fluid
  (Hayems fluid)
• Hayems Fluid
• Mercuric chloride 0.25gm
  ? Antiseptic
• Sodium chloride 0.5 gm
  ? provides correct osmotic pressure
• Sodium sulphate 2.5 gm
  ? provides correct osmotic pressure
• Distilled water
  100ml ?Solvent

30
Procedure

• Methods -Visual haemocytometric method
• -Electronic method
• Draw the blood from finger prick or
  anticoagulated blood up to 0.5 in RBC pipette
• Draw diluting fluid up to mark 101 in the RBC
  pipette
• Mix well by rotating the pipette for 2-3 minutes
• Charge the Neubauers chamber after discarding
  few drops (Diluting fluid is not mixed with
  blood)
• Allow the cells to settle down for 2 minutes.
• Count RBCs under high power in counting squares
  in the centre of the chamber.

31
Neubauer Chamber - Total RBC count -Counting
areas (Orange)
32
Calculations

• Number of cells/cu.mm No.of cells counted
• 
  -----------------------
• 
  Dilution x chamber depth x chamber area
• Cells counted N
• Dilution 1/200 (101-1/0.5)
• Chamber depth 1/10 mm
• Area counted 80 / 400 1/5 sq.mm
• N
• Total RBCs/cu.mm --------------------
• 
  1/200x1/10x1/5
• 
  20x10x5N10,000 N
• Area of central large square 1 sq.mm
• Area of a medium sized square -1/25 sq.mm
• Area of the 5square used in counting RBC is
  1/25x51/5sq.mm

33
SOURCES OF ERROR

• I.False high counts
• Inadequate wiping of excess of blood from the tip
• Blood drawn above the mark 0.5
• Diluting fluid not drawn upto mark 101
• Improper mixing
• II.False low counts
• Blood not drawn up to the mark 0.5
• Diluting fluid drawn above the mark 101
• Undue delay in counting of cells
• Faulty technique of counting

34
Normal Values

• Adult male 4.5 to 5.5 million/cu.mm
• Adult female 3.8 to 5.2 million/cu.mm
• At birth 4.0 to 6.0million/cu.mm
• Increased cell count
• Physiological -At birth
• Pathological Haemoconcentration
  Diarrhea
  Dehydration
• Cyanotic heart
  diseases
• Chronic lung
  diseases
• Polycythemia vera
• Decreased cell count
• -Anaemia
• -Haemodilution

35
Differential count

• First clean the finger with cotton and spirit.
• Puncture the pulp of the index finger.
• Wipe the first drop because it will contain
  tissue fluid.
• Keep one drop of blood in the end of a glass
  slide. Now spread the blood in to a thin smear
  with a spreader slide and keeping the spreader
  slide at 45

36
Differential count
37
Differential count

• Dry the smear and stain with Leishman stain.
• A good smear must be tongue shape, should be
  evenly spread, should occupy one third of the
  slide (it should not be too long or too short)

38
Staining procedure

• Take leishman Stain and Pour drop by drop and
  cover the smear.
• Wait for 2 minutes for fixation of smear by
  Methyl alcohol.
• Fixation means the methyl alcohol in the smear
  denatures the proteins and prevents haemolysis
  when the smear comes in contact with water and
  makes the smear stick to glass slide while
  washing with distilled water.
• Wait for two minutes, add equal amount of
  distilled water and wait for 5-10 mins.
• Wash the slide with distilled water or tap water
  try the smear and examine it under oil immersion
  objective.

39
Differential count
40
Normal values

• Neutrophils 50- 75
• Lymphocytes 25 45
• Monocytes 3-8
• Eosinophils 1-6
• Basophils 0-1

41
Haematocrit (PCV)

• Percentage of erythrocytes in a known volume of
  whole blood
• Useful Anaemia Polcythaemia
• Wintrobes tube filled with anticoagulated blood
  centrifuged at 3500rpm for 30 minutes.
• The volume occupied by the erythrocytes is
  expressed as a percentage of the total volume.

42
Centrifuge - PCV
43
Centrifuge - PCV
Plasma colour Clear -
Normal Yellow-Indirect
jaundice Pink
-Haemolysis Turbid
Hyperlipediamias
44
PCV

• Normal values
• Males 40 to 55
• Females 35 to 48
• Infants 45 to 60
• Clinical significance
• Increase PCV Polycythemia, dehydration,
  burns, and shock.
• Decrease PCV Anaemia, pregnancy.
• Sources of error
• Sample
• -Increased amount of EDTA
• -Inadequate mixing of blood
• Tubes
• -Variation in the bore size
• -Improper filling of the tube

45
Red cell indices

• MCV
• MCH
• MCHC
• Useful for Morphological classification of
  Anaemias

46
The Three Derived Indicies
MCV Hct RBC x 10 90 fl MCH Hb RBC x
10 30 pg MCHC Hb Hct x 100 33
47

• Normal values
•  MCV - 81 to 85 fl 
• MCH - 29 to 32 pg 
• MCHC - 31 to 33

48
Erythrocyte Sedimentation Rate

• The rate at which the RBCs settle from the
  plasma.
• Stage I- Rouleaux formation(Aggregation)
• RBCs come together like piles of coins, last for
  10 mts.
• Stage II- Stage of sedimentation
• RBCs settle at a constant rate-last for 30 mts.
• Stages III -Stage of packing
• Packing of RBCs occurs last for 10 mts.
• Methods Westergren Wintrobes

49
Erythrocyte Sedimentation Rate
50
Procedure

• Take 1.6ml of blood in a vial
• Add 0.4ml of anticoagulant and mix
• Draw the blood up to the 0 mark in the tube
• The tube is set up in the rack perfectly
  vertical.
• Start a stop watch or clock and take reading
• The reading corresponding to the upper layer of
  the RBC column in the pipette is taken.
• Normal
• Male 0-10mm/hr
• Female 0-20mm/hr

51
Factors Influencing ESR

• Rouleaux formation -? ESR
• Ratio of red cells to plasma
• When plasma is more - ? ESR
• When plasma is less - ? ESR
• Length of the tube-
• If length of the tube is more ESR low
• If length of the tube is short -ESR ?
• Bore of the tube
• If the bore of the tube is more ? ESR will be
  more
• If the bore of the tube is less ? ESR will be
  less
• Position of the tube
• Deviation from vertical position ? ESR
• ? Plasma globulin and Fibrinogen ? ESR
• ?Plasma globulin and Fibrinogen decreased ?ESR

52
Interpretation -

• This is not a specific test.
• An increase in ESR usually seen in chronic
  inflammatory conditions like tuberculosis,
  rheumatoid arthritis or malignancy.
• The test is useful in following disease activity
  or response to treatment once a diagnosis is
  made.

53
Platelet count

• Venous blood(Avoid finger prick blood),RBC
  Pipette , Neubauer Chamber Microscope ,
  Disposable syringe, Spirit , Cotton,Cover slip
• Diluting Fluid for Platelet Count
• 3 sodium citrate containing 1 formalin also can
  be used as diluting fluid. It, however, does not
  lyse the red blood cells.

54
Rees Ecker Diluting Fluid Composition

• S No Chemical Amount Use of the Chemical
• 1.Trisodium Citrate3.gms- Anticoagulant
• 2.Neutral Formaldehyde0.2 ml - Fixes the Platelet
• 3.Brilliant Cresyl Blue0.1 Gm - Stains the
  Platelet and Gives background
• 4.Deionised water100ml - Dissolves the chemicals
• The other diluting fluid which lyses the red
  cells is 1 (w/v) ammonium oxalate.

55
Calculations

• Platelets per cu mm (µl) (Number of platelets
  counted x Dilution) / (Volume of fluid)
• Where,
• a) Dilution 200
• b) Depth of the chamber 0.1
• c) Volume of fluid 1 x 0.1 0.1 cu mm
• d) Platelets per cu mm (µl) (Number of
  platelets x 200) / 0.1
• Number of platelets x 2000

56
Abnormalities Of Platelet Count

• Normal 1,50,000 4,00,000/cu mm (µl).
• Increased Platelet count
• Chronic Myeloid leukaemia
• Acute hemorrhage
• Decreased Platelet Count
• ITP , DIC
• Autoimmune disease like Systemic Lupus
  Erythematosis
• Viral infecton

57
Peripheral Smear

• First clean the finger with cotton and spirit.
• Puncture the pulp of the index finger.
• Wipe the first drop because it will contain
  tissue fluid.
• Keep one drop of blood in the end of a glass
  slide.
• Now spread the blood into a thin smear with a
  spreader slide and keeping the spreader slide at
  45
• Dry the smear and stain with Leishman stain.
• A good smear must be tongue shape, should be
  evenly spread, should occupy one third of the
  slide

58
PS
59
Staining procedure

• Take leishman Stain and Pour drop by drop and
  cover the smear.
• Wait for 2 minutes for fixation of smear by
  Methyl alcohol.
• Wait for two minutes, add equal amount of
  distilled water and wait for 5-10 mins.
• Wash the slide with distilled water or tap water
  try the smear and examine it under oil immersion
  objective.

60

• A Normal mature Lymphocyte is seen on the left
  compared to a segmented PMN on the right. An RBC
  is seen to be about 2/3 the size of a normal
  lymphocyte

61

• Identify the Segmented Neutrophil, Band
  Neutrophil, Lymphocyte, Monocyte, Eosinophil,
  Basophil, and Platelet in the image below

62

• The RBC's in the background appear normal. The
  important finding here is the presence of many
  PMN's. An elevated WBC count with mainly
  neutrophils suggests inflammation or infection. A
  very high WBC count (gt50,000) that is not a
  leukemia is known as a "leukemoid reaction". This
  reaction can be distinguished from malignant
  WBC's by the presence of large amounts of
  leukocyte alkaline phosphatase (LAP) in the
  normal neutrophils.

63

• The RBC's here have stacked together in long
  chains. This is known as "rouleaux formation" and
  it happens with increased serum proteins,
  particularly fibrinogen and globulins. Such long
  chains of RBC's sediment more readily. This is
  the mechanism for the sedimentation rate, which
  increases non-specifically with inflammation and
  increased "acute phase" serum proteins.

64
RBC MORPHOLOGICAL ABNORMALITIESSICKLE CELLS

• MorphologySickle shaped red cells
• Found inHb-S disease

65
RBC MORPHOLOGICAL ABNORMALITIESTEAR DROP CELLS

• MorphologyRed cells shaped like a tear drop or
  pear
• Found inBone marrow fibrosisMegaloblastic
  anaemiaIron deficiencyThalassaemia

66
RBC MORPHOLOGICAL ABNORMALITIESMALARIAL PARASITE
(P. falciparum)

• MorphologyRing form of Pl falciparum in red
  cells. Delicate rings with 1 or 2 chromatin dots.
  Often more than one ring in a red cell. Accolé
  forms are found.
• Found inMalaria

67

• The RBC's here are smaller than normal and have
  an increased zone of central pallor. This is
  indicative of a hypochromic (less hemoglobin in
  each RBC) microcytic (smaller size of each RBC)
  anemia. There is also increased anisocytosis
  (variation in size) and poikilocytosis (variation
  in shape).

68

• This Hypersegmented Neutrophil is present along
  with Macro-ovalocytes in a case of pernicious
  anemia. Compare the size of the RBC's to the
  lymphocyte at the lower left center.

69

• There are numerous fragmented RBC's seen here.
  Some of the irregular shapes appear as "helmet"
  cells. Such fragmented RBC's are known as
  "schistocytes" and they are indicative of a
  Microangiopathic hemolytic anemia (MAHA) or other
  cause for intravascular hemolysis. This finding
  is typical for disseminated intravascular
  coagulopathy (DIC).

70

• The WBC's seen here are "atypical" lymphocytes.
  They are atypical because they are larger (more
  cytoplasm) and have nucleoli in their nuclei. The
  cytoplasm tends to be indented by surrounding
  RBC's. Such atypical lymphocytes are often
  associated with Infectious Mononucleosis.

71

• Here is another example of Sickled Erythrocytes
  in a patient with Hgb SS who presented with
  severe abdominal pain in sickle crisis. The
  sickled cells are prone to stick together,
  plugging smaller vessels and leading to decreased
  blood flow with ischemia.

72
WBC MORPHOLOGICAL ABNORMALITIESTOXIC GRANULATION

• MorphologyIncreased granulation. Granulation
  more basophilic and larger than normal.
• Found inSevere bacterial infectionNon specific
  finding - seen in tissue damage of various
  types.Normal pregnancyTherapy with cytokines

73
WBC MORPHOLOGICAL ABNORMALITIESDOHLE BODIES

• MorphologySmall pale blue cytoplasmic
  inclusions, often in the periphery of the cell.
• Found inInfective and inflammatory
  statesSevere burnsTuberculosisPost
  chemotherapyPregnancy

74

• ACUTE LYMPHOBLASTIC LEUKEMIA L1

75

• These mature lymphocytes are increased markedly
  in number. They are indicative of Chronic
  Lymphocytic Leukemia, a disease most often seen
  in older adults. This disease responds poorly to
  treatment, but it is indolent.

76
ACUTE MYELOID LEUKEMIA
77

• There are numerous granulocytic forms seen here,
  including immature myeloid cells and bands. This
  condition is one of the Myeloproliferative states
  and is known as chronic myelogenous leukemia
  (CML) that is most prevalent in middle-aged
  adults. A useful test to help distinguish this
  disease is the leukocyte alkaline phosphatase
  (LAP) score, which should be low with CML and
  high with a leukemoid reaction.

78
PLATELET MORPHOLOGICAL ABNORMALITIESGIANT
PLATELETS

• MorphologyPlatelet larger than a normal red
  cell.
• Found inIncreased platelet turnoverMyeloprolife
  rative disordersMyelodysplastic disorders

79
Anaemia First Test

• RETICULOCYTE COUNT

• Fragments of nuclear material
• RNA strands which stain blue
• New Methylene Blue or Brilliant cresyl Blue

Normal Less than 2
80
Reticulocyte
No definite nucleus Reticulum of RNA Deep blue
staining Light blue cytoplasm Cell size about 10 µ
81
Reticulocytes
Leishmans
Supravital
82
Reticulocyte Production Index

• For example the RPI is calculated as follows
• Reticulocyte count 9
• Hb content 7.5 g
• Correction for Anaemia
• 9 x (7.5 15) 9 x 0.5 4.5
• Correction for increased life span
• 4.5 2 2.25
• 3. Thus, the RPI is 2.25

83
Reticulocytes
Reticulocyte
84
RETICULOCYTOSIS
85
Types of Anaemia
86
Causes of Anaemia

• Decreased production of Red Cells
• - Hypo proliferative, marrow failure
• Increased destruction of Red Cells
• - Hemolysis (decreased survival of RBC)
• Loss of Red Cells due to bleeding
• - Acute / chronic blood loss (hemorrhagic)

87
Hypoproliferative Anaemias
88
Anaemia
Hb lt 12, Hct lt 38
Hemolytic
Hypoproliferative
RPI lt 2
RPI gt 2
89
Workup Second Test

• The next step is What is the size of RBC ?
• MCV indicates the Red cell volume (size)
• Both the MCH MCHC tell Hb content of RBC
• If the RPI is 2 or less
• We are dealing with either
• Hypoproliferative anaemia (lack of raw material)
• Maturation defect with less production
• Bone marrow suppression (primary/ secondary)

90
Red Cell Size
91
Mean Cell Volume (MCV)

• RBC volume (rather) is measured by
• The Mean Cell Volume or MCV and RDW

92
Anaemia Workup - MCV
93
Anaemia Workup 3rd TestPeripheral Smear Study

• Are all RBC of the same size ?
• Are all RBC of the same normal discoid shape ?
• How is the colour (Hb content) saturation ?
• Are all the RBC of same colour/ multi coloured ?
• Are there any RBC inclusions ?
• Are intra RBC there any hemo-parasites ?
• Are leucocytes normal in number and D.C ?
• Is platelet distribution adequate ?

94
Coulter counter
95
Principle of Coulter
96

• This is Normal data from a complete blood count
  as performed on an automated instrument,
  including an automated WBC differential count.

97

• Here is data from a CBC in a person with Iron
  Deficiency Anemia. Note the low hemoglobin (HGB).
  Microcytosis is indicated by the low MCV (mean
  corpuscular volume). Hypochromia correlates here
  with the low MCH (mean corpuscular hemoglobin).

98

• The CBC here shows a markedly increased MCV,
  typical for megaloblastic anemia. The MCV can be
  mildly increased in persons recovering from blood
  loss or hemolytic anemia, because the newly
  released RBC's, the reticulocytes, are increased
  in size over normal RBC's, which decrease in size
  slightly with aging.

99

• The CBC of a patient with Microangiopathic
  Hemolytic Anemia (MAHA) demonstrates a markedly
  increased RDW (red cell distribution width) due
  to the marked variation in size and shape of the
  RBC population.

100

• Here is the CBC of a person with a severe anemia
  who underwent transfusion. This accounts for the
  dual RBC population as seen in the graph at the
  lower left. A reticulocytosis in response to
  anemia has also increased the MCV.

101
What is Anaemia ?

• Important to remember
• Anemia is a clinical sign of disease
• It is not a single disease by itself
• Need to look for the underlying cause !
• Will we ignore a fever without investigation ?
• Its diagnosis is not that simple !! Well make it
• Its very common and imp. in our practice
• Drug Rx. depends on the cause

102
Definition of Anaemia

• Decrease in the number of circulating red blood
  cell mass or Low Hb and there by O2 carrying
  capacity
• Most common hematological disorder
• Almost always a secondary disorder
• As such, critical for all practitioners to know
  how to evaluate / determine its cause / treat

103
(No Transcript)
104
Bleeding time

• Is the duration of bleeding from a standard
  puncture wound in the skin to stop bleeding.
• It is a measure of platelet count, platelet
  function, as well as integrity of vessel wall.
• This is one of the common preoperative
  investigations done before surgery.
• DUKES METHOD
• A small cut is made with a lancet in the Ear
  lobe or finger tip. Blood flow from the puncture
  is and the time taken for the bleeding to stop is
  measures with a stop watch which is taken as
  BLEEDING TIME

105
Uses Diagnosis of Bleeding Disorder. Before
Surgery. Before any tooth extraction Before
Liver or pleural or Bone marrow Puncture
Bleeding time-Dukes , IVYs Template md
106
Bleeding time

• Normal BT - 1-6 mints.
• Above 10 mints is abnormal
• Prolonged bleeding time
• Thrombocytopenia ( Decreased Platelet count)
• Defect in platelet function( Thromboasthenia)
• Drugs heparin and Aspirin
• Other method - IVYS METHOD

107
Clotting time

• The time taken for the given blood to clot is
  called Clotting time.
• It is a measure of plasma clotting factors.
• It is a screening test for coagulation disorders.
  
• Methods - 1. Capillary tube method of Wright
• 2. Lee White Method

108
Capillary tube method of Wright

• Materials RequiredCapillary tube,Spirit,Cotton
  ,LancetStop Watch
• Procedure
• First clean the pulp of the finger
• Prick the Finger
• Once bleeding starts start the stop watch and
  note the time.
• Collect the bleeding blood in capillary tubes.
• Seal the ends of the capillary tubes with
  plasticine
• Once in every 30 seconds break one capillary tube
  until a thread like clot is seen between the
  broken ends.
• Now note the time and this gives the CLOTTING
  TIME
• Normal Value for this method 5-10 mins

109
Lee White Method

• Sprit,Cotton ,Lancet,Stop Watch,Test tubes,Water
  bathDisposable syringe
• Principle- Intravenous blood when collected and
  kept out side the body in a test tube it forms a
  solid clot. The time taken to form solid clot is
  measured as CLOTTING TIME.

110
Lee Whites Method

• Take three test tubes .and put 1ml of blood in
  each test tube
• Place the test tube in a stand for 10 mins.
  (Waterbath at 37degree)
• After 10 mins take the first tube and gently tap
  every 30 seconds to test for clotting
• Once the blood in the first tube is found to be
  clotted, note the time
• Now take the second tube and start tapping every
  30 seconds, till the blood clots
• Then repeat the same with the third tube. Note
  the time.
• The time interval is taken as clotting time

111
Lee Whites Method
112
Normal (4 11 minutes)

• PROLONGED CLOTTING TIME
• Hemophilia
• VonWillibrand Disease.
• Vit K Deficiency
• DECREASED CLOTTING TIME
• Corticosteroid treatment
• Drugs like epinephrine

113
THANK YOU