HEMATOLOGIC EXAMINATIONS  Specimen

̵̵ Whole blood (EDTA)

HEMOGLOBIN DETERMINATION  Reference Range
̵̵ Male: 14 – 18 g/dL
Hemoglobin Methods ̵̵ Female: 12 – 15 g/dL
1. Copper Sulfate or Specific Gravity Method
2. Gasometric Method (Oxygen Capacity Method) Factors That Affects Hemoglobin Results
Principle: Hemoglobin will combine and 1. Age
liberate a fixed quantity of Oxygen. The blood is 2. Sex
hemolyzed with saponin and the gas is collected 3. Altitude of locality
and measured in a Van Slyke Apparatus.
3. Chemical Method (Iron Content Method) Hyperchromia – increased hemoglobin level
4. Colorimetric Methods found in:
a. Visual a. Polycythemia
 Direct Matching b. Dehydration
 Acid Hematin c. Changing from high to low altitudes
 Alkali Hematin Oligochromia – decreased hemoglobin level
b. Photoelectric found in anemias.
 Oxyhemoglobin Method – measures
plasma hemoglobin Source of Error Correction
 Cyanmethemoglobin or HiCN Drabkin’s reagent is Store in a brown bottle
Method sensitive to light or in dark place
o Standard and reference Increased WBC
method use to measure count (>20 x 109/L)
hemoglobin. Increased plt (>20 x Centrifuge the solution
o Sulfhemoglobin is not 109/L) and measure the
converted to Can cause turbidity supernatant
cyanmethemoglobin and and falsely high
cannot be measured by this result
method. Add 0.01mL of the
Lipemic sample can patient’s plasma to 5mL
Cyanmethemoglobin or HiCN Method interfere and can give of Drabkin’s reagent and
a false result use the solution as
Principle: Blood is diluted in a solution of Potassium reagent blank
ferricyanide and Potassium cyanide. The hemoglobin is HbS and HbC maybe
Dilution with distilled
oxidized to methemoglobin by the Potasssium resistant to hemolysis
water (1:2)
ferricyanide. The Potassium cyanide then converts the causing turbidity
methemoglobin to cyanmethemoglobin. The Abnormal globulin
absorbance is measured spectrophotometrically at found in multiple
Add 0.1g of Potassium
540nm. myeloma or
carbonate to Drabkin’s
Waldeström
reagent
Hb (Fe2+) + K3Fe (CN) 6 Methemoglobin (Fe 3+) macroglobulinemia
Fe3+ + KCN Cyanmethemoglobin (540nm) may precipitate

 Modified Drabkin’s Reagent Explanation of the Test

̵̵ Reagent used in hemoglobin  The hemoglobin determination test is
determination. used in:
̵̵ Pale and yellow with a pH of 7.2 ± 0.2 ̵̵ Screening for disease associated
with anemia
Composition: ̵̵ Determine the severity of
Sodium Bicarbonate 1g anemia
Potassium cyanide 52mg ̵̵ Follow the response to
Potassium ferricyanide 198mg treatment for anemia
Distilled water 1000mL ̵̵ Evaluate polycythemia
Decreased Hemoglobin Levels Methods of Hematocrit Determination
 Anemia
 Iron Deficiency, Thalassemia, Pernicious anemia Macromethods:
 Liver disease 1. Wintrobe Method – Double oxalate
 Hypothyroidism 2. Haden’s Modification Method – 1.1%
 Hemorrhage Sodium citrate
 Hemolytic anemia caused by: 3. Van Allen’s Method – 1.6% Sodium
̵̵ Transfusion of incompatible Blood citrate
 Reaction to chemical or drugs 4. Sanford – Magath Method – 1.3%
 Reaction to infectious reagent Sodium citrate
̵̵ Various Systemic Disease 5. Bray’s Method – Heparin
 Hodgkins disease
 Leukemia Micromethod:
 Lymphoma 1. Capillary puncture / Adam’s Method
 SLE ̵̵ 3 layers: plasma, buffy coat and
packed RBCs’
Increased Hemoglobin Levels ̵̵ Height of the capillary tube:
 Polycythemia 75mm
 Congestive Heart Failure ̵̵ Bore: 1.155mm
 Chronic Obstructive Pulmonary Disease ̵̵ Pricking: 2 – 3mm
̵̵ Clay: 4 – 6 mm
Variation in Hemoglobin Levels
 Occurs after transfusion, hemorrhages, burns Difference Between Micro and Macro Hct Methods
 The Hgb and Hct provide valuable information MACROMETHO MICROMETHO
in an emergency situation. D D
Method of Skin puncture
Interfering Factors of Hemoglobin Determination Blood Venipuncture or
 People living at high altitudes have increased Hb Collection venipuncture
values, as well as increased Hct and RBC. Amount of
 Excessive fluid intake causes a decreased Hgb. Larger Smaller
Blood
 Normally, the Hgb is higher in infants. Relative
 Drugs 2,000 – 2,300 10,000 rpm –
Centrifugal
 Hgb is normally decreased during pregnancy. rpm 12,000 rpm
Force
Time of Shorter (3-
ClinicalAlert of Hemoglobin Longer (30min)
Centrifugation 5min)
 The panic Hgb is less than 5.0 g/dL a conditions Simplicity of
that leads to heart failure and death. Not simple Simple
technique
 A value more than 20 g/dL leads to clogging of Can be Cannot be
the capillaries as a result of hemoconcentration. ESR
performed performed
Spilling/Leakag
HEMATOCRIT DETERMINATION (Hct) Not common Common
e
 Packed Cell Volume (PCV)
Breakage of
 It is the volume of packed RBCs that occupies a Not common Common
Buffy Coat
given volume of whole blood.
Separation of
Complete Not complete
Buffy Coat
Principle: Anticoagulated whole blood is
Cost of
centrifuged, and the total volume of the red cell Expensive Cheaper
Apparatus
mass is expressed as a percentage or a decimal
fraction.
Normal Value:
 Male: 42% - 54%
 Female: 35% - 49%
Clinical Implication Properties of Hemacytometer/Counting
1. Decreased values are indicator of: Chamber
̵̵ Anemia
̵̵ Leukemia Macroscopically:
̵̵ Lymphomas  Composed of two raised surfaces each
̵̵ Adrenal Insufficiency in the shape of a 3mm x 3mm square
̵̵ Chronic disease separated by an H – shaped moat.
̵̵ Acute and Chronic blood loss  Total ruled area: 9 sq.mm
2. Hct may or may not be reliable immediately
after even a moderate loss of blood or Microscopically:
immediate transfusion.  One large is made up of nine 1mm x
1mm squares.
Increased Hct values occur in:  Each of WBC squares is divided further
 Erythrocytosis to 16 squares.
 Polycythemia vera  The center square for RBC is subdivided
 Shock, when hemoconcentration rise into 25 smaller squares.
considerably.  The distance between each counting
surface and coverslip.
Interfering Factors in Hct Determination  Depth: 0.10mm
 High altitude  The total volume is 9 cu mm.
 Normal value vary with age and gender
 Lower value in men and women older than 60 Calculations:
y/o.
 Severe dehydration from cause falsely raises the General Formula
Hct.
cellscounted x dilutionfactor
TotalCount=
Sources of Error in Hematocrit Determination area ( mm 2 ) xdepth(0.1)
1. Speed and duration of centrifugation
̵̵ Decrease in centrifugal force will
cellscounted x dilutionfactor x 10
result in more trapped plasma in TotalCount=
between red cells. area ( mm2 )
2. Type and amount of anticoagulant
̵̵ Excess anticoagulant causes shrinkage WHITE BLOOD CELL COUNT
of cells.  Represents the number of WBCs in 1
3. Integrity in the length and diameter of the tube. liter of whole blood.
4. Errors in the sample, improper techniques in
the collection of venous and capillary blood. Formula:
5. Failure to mix the blood properly before
¿ o fcellscountedxdilutionfactor
sampling. WBCCount=
6. Leakage of blood in the case of areacounted ( mm2 ) xdepth c . f .
microhematocrit.
7. Errors in taking the reading and calculating the Variables:
result. Dilution factor = 1:20
Area counted = 4
HEMOCYTOMETRY Depth correction factor = 0.1
 The process of enumerating blood cells.
1. RBC count ¿ ofcellscountedx 20
WBC Count =
2. WBC count 4 x 0 .1
3. Platelet Count
SI Unit: no. x 109/L

Reference Range:
 3% Disodium citrate 990mL

WBC Diluting Fluid b. Hayem’s Diluting Fluid

1. 2 – 3% Glacial Acetic Acid
2. 1% HCI added with 1 drop of methyl violet or Mercuric chloride 1g
crystal violet Sodium Sulfate Anhydrous 4.4g

Criteria of Good WBC Diluting Fluid c. Gower Solution

1. Should be hypotonic. ̵̵ Prevents rouleaux formation
2. Should color/stain the nuclei of white blood
Sodium sulfate anhydrous 12.5g
cells.
Glacial acetic acid 33.3mL
Distilled water 200mL
CORRECTED WBC COUNT
 WBC count must be corrected if 5 or more
d. Toisson’s Fluid
Nucleated RBC (NRBCs) are counted on
̵̵ High specific gravity and stains the
differential count since NRBCs present in the
WBC.
sample are not lysed by the diluting fluid and
counted as WBC.
Na chloride 1g
Na Sulfate 8g
Formula:
Glycerin 30g
uncorrected WBC count x 100 Methyl violet 0.25g
CWBC= Distilled water 180mL
¿ of NRBCs per 100 WBC+ 100
e. Normal Saline Solution
̵̵ Used in emergency cases, used in the
RED BLOOD CELL COUNT
presence or rouleaux formation
autoagglutination of cells.
General Formula:
Na chloride 085g
RBC Count =¿ of cells counted x area c . f . x depthc . f . x dilution factor Distilled water 100mL
Variables: f. 3.8% Sodium Citrate
Area correction factor = 5
Depth correction factor = 10 Sodium citrate 3.8g
Dilution factor = 1:200 Distilled water 100mL
RBC count = # of cells counted x 5 x 10 x 200 OR g. Bethel’s Fluid
RBC count = # of cells counted x 10, 000

SI unit: no. x 1012/L Criteria of Good RBC Diluting Fluid

 Must be an isotonic solution
Reference Range:
 Has a good preservative
Male = 4.6 – 6.0 x 1012/L
 Does not initiate the growth of molds
Female = 4.0 – 5.4 x 1012/L
and yeast.
 With a high specific gravity
 With buffer action
RBC Diluting Fluid  Cheap and easy to prepare
a. Dacies Fluid (Formol Citrate)
̵̵ This is considered the best diluents. It RULE OF THREE
keeps for a long time and does not  The value of the hematocrit should be three times
alter the shape of the cells. the value of hemoglobin (+3).
40% formaldehyde 10mL
  This rule applies only to specimens that have ̵̵ Fish Tapeworm infestation
normocytic normochromic erythrocytes. ̵̵ Folate deficiency
̵̵ Lack of vegetable
Example:
̵̵ Alcoholism
Hgb = 12 g/dL
Hct = 36% c. Normocytic Normochromic Anemias
12 x 3 = 36
d. Anemia with Appropriate BM Response
RED BLOOD CELL INDICES ̵̵ Acute Post Hemorrhagic Anemia
 Indices define the size and Hgb content of the ̵̵ Hemolytic Anemia
RBC and consist of the:
̵̵ Mean Corpuscular Volume (MCV) e. Anemia with Impaired Marrow Response
̵̵ Mean Corpuscular Hgb ̵̵ Marrow Hypoplasia
Concentration (MCHC) ̵̵ Aplastic Anemia
̵̵ Mean Corpuscular Hemoglobin ̵̵ Marrow Infiltration
(MCH) ̵̵ Infiltration by Malignant cells,
 Help to determine specific types of anemias. Myelofibrosis
f. Decreased Erythropoietin Production
̵̵ Kidney and Liver dse
MEAN CORPUSCULAR VOLUME (MCV) ̵̵ Endocrine Deficiencies
 Indicates the mean or average volume of a red ̵̵ Malnutrition
cell. ̵̵ Anemia of Chronic dse
 Individual cell size is the best index for
classifying anemias. MEAN CORPUSCULAR HEMOGLOBIN DETERMINATION
 Index expresses the volume occupied by a single (MCHC)
erythrocyte and measures in cubic micrometers  The average concentration of hemoglobin in
(femtoliters) of the mean volume. each individual erythrocyte.
 Indicates whether the RBC size appears normal,  Measures the average conc. of hgb.
smaller than normal or larger than normal.
 Most valuable in monitoring therapy for
anemia.
Formula:
Formula:
Hct ( ) x 10
MCV = Hgb ( g /dL ) x 100
RBC ∈million MCHC=
Hct ()
Normal Value:
80 – 100 fL Normal Value:
32 – 36g/dL
SI Unit: femtoLiter (fL)
SI Unit: grams/deciliter (g/dL)
Clinical Implication
a. Microcytic Anemia (MCV 50 – 80fL) Clinical Implication
̵̵ Disorder of iron metabolism  Decreased MCHC values signify that a
̵̵ IDA unit volume of packed RBCs contain less
̵̵ Anemia of chronic disease Hgb than normal.
̵̵ Congenital hypochromic – microcytic  Hypochromic Anemia (MCHC <30g/dL)
anemia with iron overload ̵̵ IDA
̵̵ Microcytic Anemia
b. Macrocytic Anemia (MCV more than 100fL) ̵̵ Chronic Blood Loss Anemia
̵̵ Cobalamin (B12) Deficiency Hgb ( g/ dL ) x 10
MCH =
(Megaloblastic anemia – RBC ∈million
MEAN CORPUSCULAR HEMOBLOGIN
hypersegmented)
̵̵ Decreased ingestion Formula:
Normal Value:
̵̵ Competitive Parasite 27 – 33 pg

SI Unit: picrogram (pg)

 ABSOLUTE RETICULOCYTE COUNT (ARC)
Increase:  The actual number of reticulocytes in 1
 Associated with macrocytic anemia liter of whole blood.

Decrease: Formula:
 Associated with microcytic anemia
Reticulocyte () x RBC count (x 1012)
Interfering Factors
ARC =
100
 Hyperlipedemia falsely elevates the MCH.
 High heparin concentration falsely elevates Reference Range:
MCH. 25 – 75 x 109/L

Ex. Retics: 2% RBC count: 2.20 x 1012

RED CELL SIZE DISTRIBUTION WIDTH (RDW)
2 x 2.20
 Normal value: 11.5 – 14.5 CV of red cell size ARC = =44 x 109 / L
100
 Explanation of the test: Automated method of
measurement is helpful in investigation of some
hematologic disorders and in monitoring CORRECTED RETICULOCYTE COUNT
response to therapy.  In specimen with a low Hct, the
 RDW is essentially an indication of the degree percentage of reticulocytes maybe
of anisocytosis. falsely elevated because whole blood
contains fewer RBCs.
Clinical Implication  A correction factor is sued considering
 Helpful in distinguishing uncomplicated the average normal Hct to be 45%.
heterozygous Thalassemia (low MCV).
Formula:

¿
RETICULOCYTE COUNT
reticulocytes ( ) x Hct ¿
 Use to asses erythropoietic activity of the bone
Corrected RC =¿
marrow.
 Whole blood, anticoagulated with EDTA is
stained with a supravital stain such as new Reference Range:
mehtylene blue or brilliant cresyl blue or pure
azore blue.
RETICULOCYTE PRODUCTION INDEX (RPI)
Formula:
Formula:
¿ of reticulocytes
%Reticulocyte= x 100 Corrected Retics Count
1000 RBCs observed RPI=
maturation time
Reference Range:
RPI >3 adequate BM
0.5% - 1.5%
RPI <2 inadequate BM
Ex. 12 retics counted

12 x 100
% Retics = =1.2
1000
 Increased Reticulocyte Count Maturation Time of Reticulocyte (Hct)
 HA Correction Factor
Patient’s Hct Value (%) (Maturation Time in
 Lead poisoning
Days)
 Malaria
40 – 45 1
 Parasitic infections 35 – 39 1.5
 Blood intoxication 25 – 34 2
 Kala – azar 15 – 24 2.5
 Erythroblastic Anemia <15 3
 Sickle Cell Anemia ̵̵ Takes place during the 10 minutes.
 Relapsing fever (Rickettsial infxn)
 Leukemia Methods of ESR Determination
 Splenic tumor
Macromethods
Decreased Reticulocyte Count  Wintrobe – Landsberg Method
 Aplastic Anemia  Westergren Method – most common
 Acute benzol poisoning  Graphic and Cutler Method
 Chronic Infections  Linzenmeir Method
 Anaplastic crisis of HA
Micromethods
Physiologic Increase of Reticulocytes  Micro Landau Method
 Pregnancy  Smith Method
 At Birth  Hellige – Volmer Method or Crista
 Menstruation Method

ERYTHROCYTE SEDIMENTATION RATE (ESR) Reference Range for the Westergren Method
 Refers to the speed of fall of the erythrocyte to  Male: 0 – 10 mm/hr
settle down from their plasma.  Female: 0 – 15 mm/hr
 Useful in monitoring the course of an existing  Children: 0 – 13mm/hr
inflammatory disease or differentiating between Reference Range for the Wintrobe Method
similar diseases.  Male: 0 – 9mm/hr
 Female: 0 – 20mm/hr
Two Ways of Measurement
 Measuring the length of fall from the top of the Factors that Influence ESR
column of RBC in a specified period of time.  Intrinsic Factors
 Determining the time required for the red cells ̵̵ Plasma Factors
to reach a specified point. ̵̵ Red cell Factors
 Extrinsic Factors
Phases or Stages of ESR ̵̵ Mechanical Factors
1. Agglomeration Phase
̵̵ Technical Factors
̵̵ Initial period of aggregation.
̵̵ Physical Factors
̵̵ Few cells sink under gravity but the
majority form.
Factors that Increase the Rate of Fall
̵̵ Agglomerates (rouleaux) of various
 Intrinsic Factors
sizes.
o Plasma factors
̵̵ Takes place during the first 10 minutes.
̵̵ Increased Fibrinogen Conc.
2. Phase of Fast Settling
̵̵ Increased Globulin Conc.
̵̵ The agglomerates sink rapidly.
̵̵ Cholesterol
̵̵ The rate of fall depends on size.
o Red cell factors
̵̵ Takes place for about 40 minutes.
̵̵ Macrocytes
3. Final Phase of Packing
̵̵ Anemia
̵̵ The rate of settling is slow owing to
̵̵ Hemolysis
clogging of the agglomerates.
 Factor that Decrease Rate of Fall  (LE) – Lupus Erythematosus Examination
 Intrinsic Factors  Bone Marrow Examination
o Plasma factors
̵̵ Increased albumin BLOOD FILM OR SMEAR EXAMINATION
̵̵ Increased lecithin
̵̵ Defibrination Methods of Preparation of Blood Smears

 Extrinsic Factors Manual Method

̵̵ Long standing of blood since RBC 1. Wedge/Push/2 – Glass Slide Method – the
tends to be spherical. simplest and most commonly used.
̵̵ Excess dry anticoagulant
̵̵ Temp. below 200C
̵̵ Short sedimentation tube
̵̵ More blood specimen Advantages:
̵̵ Presence of blood clots ̵̵ Slides are not easily broken
̵̵ Dirty glasswares ̵̵ Easy to prepare
̵̵ Easy to label and transport
 Red Cell Factors ̵̵ Allows storages even without cover slip
̵̵ Microcytosis ̵̵ Abnormal cells can easily be found
̵̵ More red cells
̵̵ Spherocytosis Characteristics of a Good Smear
̵̵ Increased sickle cells and 1. There should be a transition from thick
Poikilocytes to thin area.
2. Smear should occupy ¾ of the length of
Increased ESR the slide.
 All collagen disease, SLE 3. Most have a smooth even surface, free
 Infection, pneumonia from waves, ridges and holes.
 Inflammatory disease 4. White blood cells should not be bunched
 Carcinoma, Lymphoma at the end or edge of smear.
 Toxemia 5. It should have a feathery edge or tail.
 Anemia
Uses of Thin Smears
Normal ESR (no increased) 1. WBC diff. count
 Sickle cell anemia 2. Stained red cell examination
 Congestive Heart Failure 3. Platelet count (indirect method)
 PK deficiency 4. Retculocyte count
5. Siderocyte count
Normal or Varied ESR 6. Malarial parasite examination
7. Thorough study of morphology of red
 Acute disease
cells.
 Musculoskeletal condition
 Cardiovascular infarction
Requirements to Produce a Proper Blood Films
 Maignant Disease
1. Use a chemically clean glass slides and
 Acute Allergy
cover glass.
2. Use of not too large or too small drop of
blood.
MORPHOLOGICAL EXAMINATION OF BLOOD CELLS
3. Work is done quickly before coagulation
of the blood.
Different Tests That Use Blood Smears
4. Proper angle and pressure of the
 Leukocyte Diff. Count
spreader.
 Morphologic Study of Normal and Abnormal
WBC, RBC and Platelets
Methods of Drying the Blood Films
 Reticulocyte Count 1. Air Drying
 Platelet Count (Indirect Method) 2. Heating in the oven for a low flame
 3. Chemical drying in ethyl alcohol
1. Wright’s Stain
Fixatives for Blood Films ̵̵ Considered polychrome stain
 Methanol component:
 Absolute Ethyl Alcohol a. Methylene blue – basic dye
 Absolute Ethyl Alcohol and Ether stains acidic cellular
 1% sol’n of Mercury Chlorideand 1% components.
Formalin b. Eosin – acid bye and stains basic
(eosinophilic) cellular
components.
̵̵ Sodium Phosphate – buffer added to
the stain and must have a pH of 6.4

Factors Affecting Thickness/Thinness of Smear 2. Giemsa Stain

3. May – Grunwald’s Stain
Factors Effects 4. Leishman’s Stain
Size of the drop of blood used 5. Jenner’s Stain
a. Large drop thick smear 6. Panoptic Stain – combination of
b. Small drop thin smear Romanowky Stain and another stain.
Angle of the spreader slide against the 7. Supravital Stain – used to stain and inspect
stationary slide living cells which have been removed from
a. Proper angle (35 – 45) good smear the body (e.g. NMB, BCB, Pure Azure Blue)
b. Increased angle thick smear
c. Decreased angle thin smear LEUKOCYTE DIFFERENTIAL COUNT
Pressure exerted when pushing the spreader  It is the enumeration and determination
against the stationary of relative proportion or percentage (%)
a. Heavy pressure thin smear of each type of leukocyte in the
peripheral or venous blood.
b. Light pressure thick smear
Speed of the spreader slide
Steps in Leukocyte Diff. Count
a. Too fast thick smear
1. Preparation of blood smear
b. Too slow thin smear
2. Staining of blood smear
3. Differentiation of leukocytes
4. Reporting of results
2. Cover Slide/Ehrlich’s Method
̵̵ Preferred for bone marrow preparation.
Ways of Scanning Smears for Diff. Count
̵̵ Even distribution of blood cells especially 1. Strip or Horizontal Method
leukocyte is observed. ̵̵ All cells in an longitudinal strip from
head to end or tail of the cells are
Disadvantages: counted.
̵̵ Cover glasses are easily broken 2. Crenellation Method
̵̵ Require chemically clean coverglass ̵̵ Cells are counted from the upper
̵̵ Difficult to prepare part of the smear, the lower par,
̵̵ Difficult to label, stain and transport then sideways, then to the upper
part until 100 cells are
3. Beacom’s Method/Cover Glass and Slide differentiated.
Method – there is even distribution of cells but 3. Exaggerated Battlement Method
yields limited blood smear. 4. Two Field Meander Method
5. Four Field Meander Method
Automated Method
 Spun Smear – prepared in Hemaspinner Shifting Process
 Smear prepared in Miniprep
Shift to the Left (immature)
Types of Stains Used for Blood Smears
 ̵̵ The presence of an increase in
younger forms of leukocytes,
̵̵ Seen in pyogenic infections.

Shift to the Right (older)

̵̵ The presence of an increase older forms
of leukocytes, MEAN CORPUSCULAR VOLUME (MCV)
̵̵ Seen in Megaloblastic Anemia,
Pernicious Anemia and in Hct ( ) x 10
MCV =
convalescence. RBC ∈million

Normal Value:
80 – 100 fL

SI Unit: femtoLiter (fL)

MEAN CORPUSCULAR HEMOGLOBIN DETERMINATION
Hemacytometer (Gen. Formula)
(MCHC)
cells counted x dilutionfactor
TotalCount= Hgb ( g /dL ) x 100
area ( mm2 ) xdepth(0.1) MCHC=
Hct ()
cellscounted x dilutionfactor x 10
TotalCount= Normal Value:
area ( mm2 )
32 – 36g/dL

WHITE BLOOD CELL COUNT SI Unit: grams/deciliter (g/dL)

¿ ofcells countedxdilutionfactor
WBCCount= MEAN CORPUSCULAR HEMOBLOG
area counted ( mm2 ) xdepth c . f .
Hgb ( g/dL ) x 10
Variables: MCH =
RBC ∈million
Dilution factor = 1:20
Area counted = 4
Depth correction factor = 0.1 Normal Value:
27 – 33 pg
¿ ofcellscountedx 20 SI Unit: picrogram (pg)
WBC Count = SI Unit: no.
4 x 0 .1
x 109/L RETICULOCYTE COUNT

¿ of reticulocytes
%Reticulocyte= x 100
CORRECTED WBC 1000 RBCs observed
uncorrected WBC count x 100
CWBC= Reference Range:
¿ of NRBCs per 100 WBC+ 100 0.5% - 1.5%
RBC Count =¿ of cells counted x area c . f . x depthc . f . x dilution factor
RBC COUNT Ex. 12 retics counted
Variables:
Area correction factor = 5
12 x 100 12
Depth correction factor = 10 Retics==Reticulocyte
% ARC () x RBC count (x 10 )
=1.2
Dilution factor = 1:200 1000
100
RBC count = # of cells counted x 5 x 10 x 200 OR ABSOLUTE RETICULOCYTE COUNT (ARC)
RBC count = # of cells counted x 10, 000 Reference Range:
25 – 75 x 109/L
SI unit: no. x 1012/L Reference Range:
Male = 4.6 – 6.0 x 1012/L Ex. Retics: 2% RBC count: 2.20 x 1012
Female = 4.0 – 5.4 x 1012/L
2 x 2.20 9
CORRECTED RETICULOCYTE COUNT

reticulocytes ( ) x Hct ( )
Corrected RC =
45

Reference Range: 2 – 3%
RETICULOCYTE PRODUCTION INDEX (RPI)

Corrected Retics Count

RPI=
maturation time

RPI >3 adequate BM

RPI <2 inadequate BM