MODULE 1 CBC Flashcards
CBC is previously know as?
Full Blood Count
CBC is consist of
Hemoglobin
Hematocrit
WBC Count
RBC Count
Differential Count
Platelet Count
RBC Indices
Factor for Hemoglobin
10
Conventional Units for Male in Hemoglobin
14-17.5 g/dL
SI units for Female Hemoglobin
123-153 g/L
SI units for Male Hemoglobin
140-175 g/L
Conventional Units for Female for Hemoglobin
12.3-15.3 g/dL
Principle of Hemoglobin
Used to diagnose and follow treatment of anemia
Hemoglobin is INCREASED in:
Polycythemia vera
Morning
Strenuous Exercise
High Altitudes
Hemoglobin is DECREASED in:
Anemia
Hemoglobin is SLIHTLY LOWER in:
Patient lying down
Cyanmethhemoglobin method is also know as
Hemiglobincyanide Method
Principle of Cyanmethemoglobin
Reference method for hemoglobin
Measure all forms of hemoglobin except SULFHEMOGLOBIN
Drabkin’s Reagent Consist of:
Potassium Ferricyanide
Potassium Cyanide
Dihydrogen Potassium Phosphate
Nonionic Detergent
Distilled Water
Converts HEMOGLOBIN into METHEMOGLOBIN
Potassium Ferricyanide
Convers METHEMOGLOBIN into CYANMETHEMOGLOBIN
Potassium Cyanide
Shortens conversion time from 10-15 mins. to 3 mins.
Dihydrogen Potassium Phosphate
Enhances LYSIS of RBC’s
Nonionic Detergent
Example of NONIONIC DETERGENT
Steroz
Triton X
Drabkin’s reagent should be
Clear and pale yellow
Very poisonous and must be kept in a locked cabinet at all times when not in use.
Potassium Cyanide
Sources of Error in Hemoglobin Dtermination
Lipemia
Increased WBC’s and Platelets
HbS and HbC
Increased Globulins
Overanticoagulation
Correction need for LIPEMIA
corrected using patient blank
Correction need for Increased WBC’s and Platelets
Corrected by centrifuging test mixture and testing hemoglobin on the supernatant fluid
Correction need for HbS and HbC
Dilute hemoglobin with distilled water
Correction need for Increased globulins
Use of Dihydrogen Potassium Phosphate
Principle of HEMATOCRIT
Term Packed Cell Volume; measure how much is your blood consist or RBC
HEMATOCRIT term used to describe the method.
Relationship of Hb and Hct in Ratio
1:3
The relationship of Hb and Hct in Ratio may vary in
Cause of anemia and the effect on the RBC Indices especially in MCV.
Factor in Hct
0.01
SI units for Male Hct
0.415-0.504 volume fraction
Conventional Units for Male Hct
41.5-50.4%
Conventional Units for Female Hct
35.9-44.6%
SI units for Female Hct
0.359-0.446 volume fraction
Reference manual method for Microhematocrit
Spun Microhematocrit
Microhematocrit tube long, bore and holds blood..
75mm
1mm
0.05 mL
Anticoagulant for RED Capillary Tube
Sodium Heparin
Anticoagulant for Blue Capillary Tube
Plain
Anticoagulant for Green Capillary Tube
Ammonium Heparin
Anticoagulant for ORANGE Capillary Tube
Lithium Heparin
Clay plug should be ___mm long
4-6 mm long
Layers of Blood after cnetrifugation
Fatty Layer
Plasma
Buffy Coat
Packed RBC’s
Sealing Clay
Color of Bead in Thoma Pipette for WBC
White
Color of Bead in Thoma Pipette for RBC
Red
Volume of the Bulb for Thoma Pipette for RBC
100
Volume of the Bulb for Thoma Pipette for WBC
10
Dilution Range for Thoma Pipette for RBC
1:100-1:1000`
Dilution Range for Thoma Pipette for WBC
1:10-1:100
Outstanding Marks for Thoma Pipette for RBC
0.5, 1, 101
Outstanding Marks for Thoma Pipette for WBC
0.5, 1, 11
Factor in RBC Count and WBC Count
10^6
SI Units for Female in RBC Count
4.5-5.1 X 10^12/L
SI Units for Male in RBC Count
4.5-5.9 X 10^12/L
Conventional Units for Male in RBC Count
4.5-5.9 X 10^6 u/L
u/L- units per liter
Conventional Units for Female in RBC Count
4.5-5.1 X 10^6 u/L
Hct result should be agree with
+ - 0.02 L/L
L/L- Liter of cells per liter of blood
RBC is HIGHER in
Newborns
RBC is SLIGHTLY DECREASE in
Childhood/adolescence and after 50 yrs of age
RBC is INCREASED in
Polycythemia
Dehydration
High Altitudes
RBC is DECREASED in
Anemia
Procedure of Manual RBC Count
- Draw blood up to 0.5 mark of thoma pipette, and add the diluting fluid for 101 mark
- Shake the pipette in a mechanical shaker for 3 mins.
- Discard the first 4 drops from the pipette and fill the counting chamber.
- Count RBC’s in 5 Small central squares.
- Calculate the RBC count
- Total no. of RBC’s Counted X 50
Dilution of RBC Count
1:200
Dilution for Polycythemia
1:300
Dilution for Severe Anemia
1:100
Principle of RBC Diluting Fluid
Should be isotonic to prevent lysis of RBC’s
Examples of RBC’s Diluting Fluid
Hayem’s
NSS
Dacie’s (Formol-Citrate)
Toisson’s
Gower’s
Bethel’s
3.8% Na Citrate
Conventional Units for WBC Count
4.4-11 X 10^3/uL
/uL- Microliters
SI Units for WBC Count
4.4-11.3 X 10^9/L
WBC Count is Higher in
Newborns
1 yr old
Adult
WBC in Newborns
10-30 X 10^9/L
WBC in 1 year old
6-17 X 10^9/L
WBC in Adult
4-11 X 10^9/L
Diurnal Variation of WBC
Highest in AFTERNOON
Lowest in MORNING
Anticoagulant used for WBC
EDTA
Unsatisfactory Anticoagulant for WBC
Heparin
Principle of WBC Diluting Fluid
Lyses the RBC so that they will not obscure the WBC
Principle of WBC Diluting Fluid
Lyses the RBC so that they will not obscure the WBC
Example of WBC Diluting Fluid
1% Ammonium Oxalate
2-3% Glacial Acetic Acid with Gentian Violet
1% Hydrochloric Acid (IMH Used)
Turk’s Solution (Glacial Acetic Acid & methyl Violet)
Procedure in WBC Count
1.Draw blood exactly 0.5 mark and dilute up to 11 mark with diluting fluid
2. Mix the pipette in the mechanical shaker for 3 Mins.
3. Discard first 4 drops of the mixture and fill the hemocytometer
4. Count the 4 Corner Square using LPO
5. Calculate the WBC count by multiplying the # of WBCs Counted in 4 Squares by 50
Cleaning Solution for the Neubauer Chamber
95% Alcohol
Cells that are not lysed by the WBC Diluting Fluid
Sickle Cells
Target Cells
Wedge Blood Smear is also known as
Spreader Slide
Push Smear
2 Slide method
Position of spreader slide
30-45 degree angle
Extremely Thick Smears are caused by:
Decreased pressure
Increased angle of spreader slide
Increased size of blood
Increased speed of spreader
Extremely Thin Smears are caused by:
Increased pressure
Decreased angle of spreader slide
Decrease size of blood
Decrease speed of spreader
Other types of Blood Smears
Cover Slide Method
Automation Blood Films
Buffy Coat Smear
Thick and Thin Blood Film
Principle of Cover Slide Method
2 cover slide are used
utilized extensively for BM exams
Procedure for Cover Slide Method
- Small drop of blood in one cover slide and place another cover slip on the top (8 pointed star overlapping)
- If spread is complete, pull 2 slide apart.
- Air dry and already ready for staining.
Principle of Wedge (Push) Type
Stimulate the manual spreader technique
Principle of Buffy Coat Smear
Centrifuge Blood in wintrobe tube for 15 mins. at 15000g or 6 mins. at 1000g and aspirate buffy coat.
Easier to locate bacteria or parasite
What is a NONVITAL POLYCHROME STAIN
Produce multiple colors when applied to cellular elements
Ex. of NONVITAL POLYCHROME STAIN
Romanowsky Stains
What is a NONVITAL MONOCHROME STAIN
Stain specific cellular components
Ex. of NONVITAL MONOCHROME STAIN
Prussian Blue
Principle of Prussian Blue
Visualizes iron granules in RBC
What is a SUPRAVITAL STAIN
Stain specific cellular components in the living state. No fixative are used in the staining process.
Ex. of SUPRAVITAL STAIN
New Methylene Blue
Principle of NEW METHYLENE BLUE
Precipitate RNA in reticulocytes
Basic Dye
+ Charge
stains acidic compound of the cell (Nucleus blue/purple)
Principle of Romanowsky Stain
Routinely used to stain peripheral blood and BM Smears
considered to be polychromatic stain
basically contain methylene blue and eosin
Eosin
Acidic Dye
- Charge
Stains Basic Compound of the Cell (cytoplasm oramge/pink)
Eosin
Acidic Dye
- Charge
Stains Basic Compound of the Cell (cytoplasm oramge/pink)
Principle of DIFFERENTIAL COUNT
Determine the relative number of each type of WBC and at the same time a study of RBC, WBC and platelet morphology is performed.
3 part differential count
granulocyte, lymphocyte and monocytes
5 part differential count
Neutrophils
Eosinophils
Basophils
Lymphocytes
Monocytes
Relative Value of Neutrophil Segmented
50-70%
Absolute Value of Neutrophil Segmented
1.8-7.8 X 10^9/L
Relative Value of Neutrophil Band
0-5%
Absolute Value of Neutrophil Band
0-0.70 X 10^9/L
Relative Value of Eosinophil
1-3%
Relative Value of Basophil
0-2%
Relative Value of Lymphocytes
18-42%
Relative Value of Monocytes
2-11%
Absolute Value of Eosinophil
0-0.45 X 10^9/L
Absolute Value of Basophil
0-0.20 X 10^9/L
Absolute Value of Lymphocytes
1.0-4.8 X 10^9/L
Absolute Value of Monocytes
0-0.80 X 10^9/L
Counting Methods
Cross Sectional or Crenellation
Longitudinal
Battlement
Principle of Cross Sectional or Crenellation
WBCs are counted consecutive fields as the blood film is moved from side to side
Principle of Longitudinal
Ideal method if the smear is thin enough
Counted in consecutive fields from tail toward the head of the smear
Principle of Battlement
Uses a pattern of consecutive fields beginning near the tail on a horizontal edge.
Principle of RBC Indices
Useful in the morphologic characterization of anemia
Calculations introduced by Wintrobe
Conventional Unit of MCV
80-96 um3
um3-one cubic microliter
Conventional Unit of MCH
27.5-33.2 pg
picogram
Conventional Unit of MCHC
33.4-35.5%
Factor of MCV and MCH
1
Factor for MCHC
0.01
SI Units for MCHC
0.334-0.355
SI Units for MCV
80-96 fL
femtoliter
SI Units for MCH
27.5-33.2 pg
MCV
Mean Corpuscular Value
MCV= Hct/RBC X 10
Indicate average volume of RBC
Interpretation of MCV
Macrocytic RBC >96 fL
Normocytic RBC 80-96 fL
Microcytic RBC <80 fL
MCH
Mean Corpuscular Hemoglobin
MCH= Hb/RBC X 10
Average content of Hb in RBC
Less valuable than MCV and MCHC
Interpretation MCH
INCREASED in MACROcytic Anemia
DECREASED in MICROcytuc Anemia and Hypoochromic Anemia
MCHC
Mean Corpuscular Hemoglobin Concentration
MCHC= Hb/Hct X 100
Average concentration of Hb in each individual RBC in g/dL
Interpretaion of MCHC
Hyperchromia >35.5% (sperocytes)
Normochromia 33.4-35.5%
Hypochromis <33.4% (IDA and Thalassemia)
RDW
Red Blood Cell Distribution Width
RDW= SD of MCV/Mean of MCV X 100
Indicates degree of anisocytosis (variation of cell size)
Determine from the RBC Histogram
RDW Normal Value
12-17%