MTAP W2 (Hematology W10: Lab Evaluation) Flashcards
CBC IS ALSO KNOWN AS
HEMOGRAM
INCLUDES
1. ENUMERATION OF CELLULAR ELEMENTS
2. QUANTITATION OF HGB
3. STATISTICAL ANALYSES OF CELL APPEARANCES
CBC/HEMOGRAM
PREVENTS PLATELETS FROM CLUMPING ON GLASS SLIDE MAKING IT MORE ACCURATE DURING FILM EVALUATION
EDTA
SPECIMEN OF CHOUCE FOR EVALUATION OF BLOOD CELL MORPHOLOGY
ANTICOAGULANT-FREE BLOOD
PERIPHERAL BLOOD FILM
EDTA BLOOD (PURPLE TOP)
MADE WITHIN – OF DRAWING THE SPECIMEN- HIGH QUALITY FILMS
2-3 HOURS
ADVANTAGES
1. MULTIPLE SLIDES
2. PREP AT LATER TIME
3. AVOIDS PLT CLUMPING
EDTA
PREFERRED EDTA TUBE WHEN MAKING PERIPHERAL BLOOD FILM
IT MIXES MORE EASILY WITH BLOOD
TRIPOTASSIUM EDTA/LIQUID FORM
PERIPHERAL BLOOD FILM
EDTA TUBES THAT REMAIN FOR MORE THAN -
BLOOD CELL ARTIFACTS:
1. VACUOLATED NEUTROPHILS
2. ECHINOCYTIC RBCS
3. NECROBIOTIC LEUKOCYTES
4. SPHEROCYTES
5 HOURS
PERIPHERAL BLOOD FILM
EDTA LEFT FOR 5 HOURS
BLOOD CELL ARTIFACTS (4)
- VACUOLATED NEUTROPHILS
- ECHINOCYTIC RBCS
- NECROBIOTIC LEUKOCYTES
- SPHEROCYTES
VENS
PERIPHERAL BLOOD FILM
CAUSE PLATELET SATELLITOSIS
EDTA SPECIMEN
PERIPHERAL BLOOD FILM
PLATELETS SURROUND OR ADHERE TO THE NEUTROPHILS
PLATELET SATELLITOSIS
PERIPHERAL BLOOD FILM
PLATELET SATELLITOSIS:
FALSE DECREASE DUE TO ADHERANCE TO NEUTROPHIL
PSEUDOTHROMBOCYTOPENIA
PERIPHERAL BLOOD FILM
PLATELET SATELLITOSIS:
FALSE INCREASE OF WBC, CLUMPS WILL BE READ AS WBC DUE TO ITS SIZE
PSEUDOLEUKOCYTOSIS
PLATELET SPECIFIC AUTOANTIBODIES THAT REACH BEST AT RT ARE ONE OF THE MECHANISMS KNOWN TO CAUSE THIS PHENOMENON
PLATELET SATELLITOSIS
PERIPHERAL BLOOD FILM
PLATELET SATELLITOSIS:
CORRECTION: RECOLLECT BLOOD SAMPLE USING -
3.2% NA CITRATE (BLUE TOP)
PERIPHERAL BLOOD FILM
PLATELET SATELLITOSIS:
CORRECTION: RECOLLECT BLOOD SAMPLE USING 3.2 NA CITRATE
ADJUSTMENT MUST MULTIPLY TO -
PLATELET COUNT X 1.1
PERIPHERAL BLOOD FILM
ADVANTAGE:
1. MADE AT PX SIDE
2. SOME ARTIFACTS MAY BE PREVENTED
ANTICOAGULANT-FREE BLOOD
METHODS OF BLOOD FILM PREPARATION
EASIEST, MOST CONVENIENT, COMMONLY USED
MANUAL/PUSH-WEDGE METHOD
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
USES SLIDE AS PUSHER/SPREADER
FILM SLIDE: 3X1 INCH (75 X25 MM)
ANGLE OF PUSHER/SLIDE: -
30-45 DEGREE
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
BLOOD IS DROPPED AT ABOUT - FROM END OR FROSTED
0.25/ 1/4 INCH
1 CM
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
DROP OF BLOOD: -
2-3MM
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
DROP OF BLOOD: 2-3MM
LARGE AMOUNT OF BLOOD= - SMEAR
THICKER/LONGER
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
DROP OF BLOOD: 2-3MM
SMALL AMOUNT OF BLOOD= -SMEAR
THINNER/SHORT
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
SPEED OF SPREADER:
TOO FAST= - SMEAR
THICKER
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
SPEED OF SPREADER:
TOO SLOW= - SMEAR
THINNER
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
HEMATOCRIT OF PX:
TOO HIGH (>60%) ANGLE SHOULD BE LOWERED AS LOW AS -
25 DEGREE
METHODS OF BLOOD FILM PREPARATION
MANUAL WEDGE TECHNIQUE aka PUSH-WEDGE METHOD
HEMATOCRIT OF PX:
TOO LOW (>45%) ANGLE SHOULD BE RAISED -
> 45 DEGREES
METHODS OF BLOOD FILM PREPARATION
TOO THICK SMEARS
PRESSURE: -
LOW
METHODS OF BLOOD FILM PREPARATION
TOO THICK SMEARS
ANGLE: -
HIGH
METHODS OF BLOOD FILM PREPARATION
TOO THICK SMEARS
SPEED: -
FAST
METHODS OF BLOOD FILM PREPARATION
TOO THICK SMEARS
SIZE OF BLOOD: -
LARGE
PERIPHERAL BLOOD FILM
COVERSLIP METHOD OF BLOOD FILM PREPARATION
- METHOD: GLASS SLIDE + COVERSLIP
BEACOM
PERIPHERAL BLOOD FILM
COVERSLIP METHOD OF BLOOD FILM PREPARATION
- METHOD: 2 COVER SLIP METHOD
EHRLICH METHOD
PERIPHERAL BLOOD FILM
FEATURES OF A WELL-MADE WEDGE PBS
FILM IS - LENGTH OF SLIDE
2/3-3/4
PERIPHERAL BLOOD FILM
FEATURES OF A WELL-MADE WEDGE PBS
THE FILM SHOULD BE - SHAPED NOT BULLET
FINGER
PERIPHERAL BLOOD FILM
FEATURES OF A WELL-MADE WEDGE PBS
SLIDE HELD UP TO THE LIGHT THIN PORTION MUST BE HAVE A - APPEARANCE
RAINBOW
PERIPHERAL BLOOD FILM
FILM THAT IS - (COLOR) THAN NORMAL MAY INDICATE THAT THE PX HAS
- INCREASED BLOOD PROTEINS
- PLASMA CELL MYELOMA
- ROULEAUX
BLUER
PERIPHERAL BLOOD FILM
A - APPEARANCE TO THE FILM MAY INDICATE RBC AGGLUTINATION FOUND IN COLD HEMAGGLUTIN DISEASES
GRAINY
PERIPHERAL BLOOD FILM
- ALL OVER THE FILM INDICATES INCREASED LIPID LEVELS
HOLES
PERIPHERAL BLOOD FILM
SLIDES STAINED AFTER - OR LONGER TURN OUT TOO BLUE
SHOULD ONLY BE STORED FOR -
1 WEEK
PERIPHERAL BLOOD FILM
FILMS PRODUCED EVERY 30SEC
SYSMEX SP10
PERIPHERAL BLOOD FILM
AUTOMATED SLIDE STAINERS
MIDAS AND HEMA TEK
PERIPHERAL BLOOD FILM
DEMONSTRATES LE CELLS
CONCENTRATES THE NUCLEATED CELLS PRESENT
BUFFY COAT SMEAR
PERIPHERAL BLOOD FILM
USES BUFFY COAT SMEAR ON PATIENTS WITH EBC COUNT
<1X10^9/L
PERIPHERAL BLOOD FILM
-BLOOD SMEAR USED FOR SCREENING OF BLOOD PARASIRES
THICK SMEARS
PERIPHERAL BLOOD FILM
- STAIN USED FOR BM AND PBS
WRIGHT/ GIEMSA STAIN
PERIPHERAL BLOOD FILM
FIXATIVE: -
METHANOL
PERIPHERAL BLOOD FILM
ACTUAL STAINING DOES NOT OCCUR UNTIL - IS ADDED
BUFFER
PERIPHERAL BLOOD FILM
-: STAINS ACIDIC PARTS OF CELL SUCH AS RNA
METHYLENE BLUE
PERIPHERAL BLOOD FILM
-: STAINS BASIC PART OF CELL
EOSIN
PERIPHERAL BLOOD FILM
OXIDIZE METHYLENE BLUE AND EOSIN
STAINS NEUTRAL COMPONENT
THIZAINE-EOSINATE COMPLEX
PERIPHERAL BLOOD FILM
BUFFER IS ADDED TO STAIN SHOULD BE -M
0.05M SODIUM PHOSPHATE
PERIPHERAL BLOOD FILM
BUFFER ADDED TO STAIN FOR ATLEAST 24 HOURS
AGED DISTILLED WATER
MOST IMPORTANT PART OF STAINING
PH 6.4-6.8
PERIPHERAL BLOOD FILM
PH FOR MALARIAL PARASITES
7.2
WELL STAINED PERIPHERAL BLOOD FILM
MACRO: SHOULD BE (COLOR)
PINK TO PURPLE
WELL STAINED PERIPHERAL BLOOD FILM
MICRO: RBC SHOULD APPEAR
ORANGE-SALMON PINK
WELL STAINED PERIPHERAL BLOOD FILM
MICRO: WBC SHOULD BE
PURPLE TO BLUE
WELL STAINED PERIPHERAL BLOOD FILM
MICRO: CYTOPLASM OF NEUTROPHILS SHOULD BE
PINK TO TAN WITH VIOLET GRANULES
WELL STAINED PERIPHERAL BLOOD FILM
MICRO: EOSINOPHILS SHOULD HAVE - GRANULES
BRIGHT ORANGE
WELL STAINED PERIPHERAL BLOOD FILM
EXCESSIVELY - (COLOR) STAIN
- THICK
- PROLONGED STAINING
- INADEQUATE WASHING
- TOO ALKALINE
BLUE
PERIPHERAL BLOOD FILM
STAINING PROBLEM:
FAILURE TO HOLD THE SLIDE -
HORIZONTALLY
COMPLETE BLOOD COUNT
PERFORMED USING A - OR -
HEMACYTOMETER/ COUNTING CHAMBER
MOST COMMONLY USED HEMACYTOMETER
LEVY WITH NEUBAUER RULING
LEVY WITH NEUBAUER RULING
COMPOSED OF -MMX-MM SQUARE COUNTING AREA SEPARATED BY AN H SHAPED MOAT
3X3MM
TOTAL= 9MM2
LEVY WITH NEUBAUER RULING
TOTAL VOLUMR OF ONE ENTIRE GRID OF HEMACYTOMETER IS
0.9MM3
LEVY WITH NEUBAUER RULING
- EACH SECONDARY SQUARE EXCEPT CENTER
16 SQUARES
LEVY WITH NEUBAUER RULING
CENTER SECONDARY SQUARES -
25 SQUARES
LEVY WITH NEUBAUER RULING
READING OF PLATELETS
CENTER SQUARE
25 SECONDARY SQUARES
LEVY WITH NEUBAUER RULING
READING OF RBC
CENTER SQUARE
ONLY 4 CORNER AND CENTER OF SECONDARY SQUARES
LEVY WITH NEUBAUER RULING
READING OF WBC
4 CORNER SQUARES
16 SECONDARY SQUARES
LEVY WITH NEUBAUER RULING
CORNER SQUARES
16 SECONDARY SQUARES EACH WITH AREA OF -
0.0625 MM2
LEVY WITH NEUBAUER RULING
CENTER SQUARE
25 SECONDARY SQUARES EACH WITH AREA OF -
0.04MM2
HEMACYTOMETER
DEPTH: -
0.1MM
HEMACYTOMETER
DEPTH FACTOR: -
1/10 OR 10
HEMACYTOMETER
DISTANCE BETWEEN THE BOTTOM OF THE COVERSLIP AND SURFACE OF COUNTING AREA
DEPTH FACTOR
DILUTING PIPETTES
COMPOSED OF STEM DIVIDED INTO - EQUAL PARTS WITH BEAD THAT AIDS IN MIXING THE DILUENT
10 EQUAL PARTS
DILUTING PIPETTES
IN CALCULATING DILUTION, ONLY THE VOLUME CONTAINED IN THE - IS CONSIDERED
BULB
DILUTING PIPETTES
RBC PIPETTE: - MARK/ TOTAL UNITS
101 MARK
DILUTING PIPETTES
WBC PIPETTE: - MARK/ TOTAL UNITS
11 MARK
DILUTING PIPETTES
RBC PIPETTE
BULB: - UNIT
100 UNIT
DILUTING PIPETTES
RBC PIPETTE
STEM: - UNIT
1 UNIT
DILUTING PIPETTES
RBC PIPETTE
BLOOD DRAWN ON 0.5 MARK
DILUTION: -
1:200
DILUTING PIPETTES
RBC PIPETTE
BLOOD DRAWN ON 1.0 MARK
DILUTION: -
1:100
DILUTING PIPETTES
RBC PIPETTE
DILUENT IS DRAWN TO THE - MARK
101 MARK
DILUTING PIPETTES
WBC PIPETTE
BLOOD DRAWN ON 0.5 MARK
DILUTION: -
1:20
DILUTING PIPETTES
WBC PIPETTE
BLOOD DRAWN ON 1.0MARK
DILUTION: -
1:10
DILUTING PIPETTES
WBC PIPETTE
DILUENT IS DRAWN TO THE - MARK
11 MARK
DILUTING PIPETTES
WBC PIPETTE
VOLUME OF THE BULB
10
GENERAL FORMULA FOR MANUAL CELL COUNTS
TC= CC X DF/ AREA X 0.1
GENERAL FORMULA FOR MANUAL CELL COUNTS
THE DIFFERENCE BETWEEN THE TOTAL CELLS COUNTED ON EACH SIDE SHOULD BE -
<10%
MANUAL CELL COUNT
CONVENTIONAL UNIT
CELLS/UL
CELLS/MM3
MANUAL CELL COUNT
SI UNIT
CELLS/L
MANUAL CELL COUNT
REPORTING RBC
CONVENTIONAL UNIT
X10^6/MM3
X10^6/UL
MANUAL CELL COUNT
REPORTING WBC/PLPATELETS
CONVENTIONAL UNIT
X10^3/MM3
X10^3/UL
MANUAL CELL COUNT
REPORTING RBC
SI UNIT
X10^12/L
MANUAL CELL COUNT
REPORTING WBC/PLATELETS
SI UNIT
X10^9/L
WBC
DILUTION: 1:20
OBJECTIVE: 10X (LPO)
AREA COUNTED: -
4MM2
WBC
DILUTION: 1:100
OBJECTIVE:10X (LPO)
AREA COUNTED: -
9MM2
RBC
DILUTION: 1:100
OBJECTIVE: 40X (HPO)
AREA COUNTED: -
0.2MM2/ 5 SMALL SQAURES OF CENTER SQUARE
PLATELETS
DILUTION: 1:100
OBJECTIVE: 40X PHASE CONTRAST
AREA COUNTED: -
1MM2
WBC DILUTING FLUIDS
- 1% AMMONIUM OXALATE
- 1% HYDROCHLORIC ACID
- 3% ACETIC ACID
RBC DILUTING FLUID
ISOTONIC SALINE
PLATELETS DILUTING FLUID
1% AMMONIUM OXALATE
MANUAL RBC COUNT ARE RARELY PERFORMED
(2) CONCENTRATIONS ARE MORE DESIRABLE WHEN AUTOMATION IS NOT AVAILABLE
- MICROHEMATOCRIT
- HEMOGLOBIN
NORMAL RBC COUNT
MALE: -/L
4.20-6.00
X10^12/L
NORMAL RBC COUNT
FEMALE: -/L
3.80-5.20
X10^12/L
MANUAL RBC COUNT
AREA OF COUNTING CHAMBER USED: 5 CENTER SQUARES
EACH SQUARE: -
0.04MM
MANUAL RBC COUNT
AREA OF COUNTING CHAMBER USED: 5 CENTER SQUARES
5 SQUARES: -
0.2MM2
MANUAL RBC COUNT
MICROSCOPE OBJECTIVE: -
40X/ HPO
MANUAL RBC COUNT
USUAL DILUTION: -
1:100
MANUAL RBC COUNT
RBC DILUTING FLUIDS SHOULD BE - TO FACILITATE COUNTING AND PREVENT LYSIS OF RBCS
ISOTONIC
MANUAL RBC COUNT
BEST DILUTING FLUID FOR RBC
DACIES/FORMOL CITRATE
MANUAL RBC COUNT
RBC DILUTING FLUIDS (7)
1.DACIES/FORMOL CITRATE
2.3.8% SODIUM CITRATE
3.NSS
4.BETTHELL’S
5.HAYEM’S
6.TOISSON’S
7.GOWER’S
RBC PARAMETERS REPORTING
RBC COUNT
RBCS
X10^12/L
RBC PARAMETERS REPORTING
HGB
g/dL
RBC PARAMETERS REPORTING
HCT
% or L/L
RBC PARAMETERS REPORTING
MCV
fL
RBC PARAMETERS REPORTING
MCH
pg
RBC PARAMETERS REPORTING
MCHC
g/dL
RBC PARAMETERS REPORTING
RDW
%
AN IMPORTANT TOOL TO ASSESS BM’S ABILITY TO INCREASE RBC PRODUCTION IN RESPONSE TO ANEMIA
RETICULOCYTE COUNT
ERYTHROCYTE MATURATION SERIES
LAST STAGE CAPABLE OF HGB SYNTHESIS
RETICULOCYTE
YOUNG RBC THAT LACKS NUCLEUS BUT CONTAINS RESIDUAL RNA TO COMPLETE PRODUCTION OF HGB
RETICULOCYTE
RETICULOCYTE
SPENDS - IN THE BM
2 DAYS
RETICULOCYTE
SPENDS - IN THE PERIPHERAL BLOOD BEFORE DEVELOPING INTO A MATURE RBC
1 DAY
RETICULOCYTE COUNT
ADULT: -
0.5-1.5%
RETICULOCYTE COUNT
NEWBORN: -
2-6%
RETICULOCYTE
STAINED WITH -
SUPRAVITAL STAIN- NEW METHYLENE BLUE
RETICULOCYTE COUNT
RATIO OF BLOOD AND STAIN
1:1
RETICULOCYTE COUNT
INCUBATE AT RT FOR -
3-10MIN
RETICULOCYTE COUNT
COUNT - RBCS UNDER - LENS
1000, OIO
RETICULOCYTE COUNT
FOR ACCURACY, HAVE ANOTHER LABORATORIAN TO COUNT SHOULD AGREE WITHIN -
20%
RETICULOCYTE COUNT
FORMULA
RETIC%= NO.OF RETIC X 100/ 1000 (RBCS COUNTED)
THE ACTUAL NUMBER OF RETICULOCYTES IN 1 LITER OR 1 MICROLITER OF BLOOD
ABSOLUTE RETICULOCYTE COUNT
ABSOLUTE RETICULOCYTE COUNT
REFERENCE INTERVAL
20X109/L
115X109/L
ABSOLUTE RETICULOCYTE COUNT
FORMULA
ARC= RETICS X RBC COUNT/100
IN SPX WITH LOW HEMATOCRIT, RETICULOCYTES MAY BE FALSELY -
ELEVATED
CORRECTED RETICULOCYTE COUNT
AVERAGE NORMAL HCT -%
45%
CORRECTED RETICULOCYTE COUNT
FORMULA
CRC= RETIC X (PX HCT/45)
CORRECTED RETICULOCYTE COUNT
PX WITH HEMATOCRIT OF 35% SHOULD HAVE AN ALEVATED RETICULOCYTE COUNT OF - TO -%
35%=MILD ANEMIA
2-3%
CORRECTED RETICULOCYTE COUNT
PX WITH HEMATOCRIT OF LESS THAN 25% COUNT SHOULD INCREASE TO - TO -%
<25%= MODERATE ANEMIA
3-5%
GENERAL INDICATOR OF THE RATE OF ERYTHROCYTE PRODUCTION INCREASE ABOVE NORMAL IN ANEMIAS
RETICULOCYTE PRODUCTION INDEX
RETICS THAT ARE RELEASE FROM BM PREMATURELY ARE CALLED -
USUALLY TO COMPENSATE ANEMIA
SHIFT RETICULOCYTES
SHIFT RETICS TAKES - TO - DAYS TO LOSE THEIR RETICULA
2-3 DAYS
RETICULOCYTE PRODUCTION INDEX
FORMULA
RPI= CORRECTED RETIC COUNT/MATURATION TIME
THE PX - IS USED TO DETERMINE APPROPRIATE CORRECTION FACTOR
HEMATOCRIT
RETICULOCYTE PRODUCTION INDEX
PX HEMATOCRIT VALUE %
40-45
CORRECTION FACTOR/MATURATION TIME/DAYS
1
RETICULOCYTE PRODUCTION INDEX
PX HEMATOCRIT VALUE %
35-39
CORRECTION FACTOR/MATURATION TIME/DAYS
1.5
RETICULOCYTE PRODUCTION INDEX
PX HEMATOCRIT VALUE %
25-34
CORRECTION FACTOR/MATURATION TIME/DAYS
2
RETICULOCYTE PRODUCTION INDEX
PX HEMATOCRIT VALUE %
15-24
CORRECTION FACTOR/MATURATION TIME/DAYS
2.5
RETICULOCYTE PRODUCTION INDEX
PX HEMATOCRIT VALUE %
<15
CORRECTION FACTOR/MATURATION TIME/DAYS
3
RETICULOCYTE PRODUCTION INDEX
ADEQUATE BM RESPONSE INDICATED BY AN RPI -
> 3
RETICULOCYTE PRODUCTION INDEX
INADEQUATE ERYTHROPOIETIC RESPONSE IS SEEN WHEN THE RPI -
<2
RETICULOCYTE COUNT
DESIGNED TO REDUCE LABOR INTESIVE PROCESS
DISC INSERTED INTO THE EYEPIECE OF MICROSCOPE
MILLER DISK
RETICULOCYTE COUNT
2 SQUARES SMALLER SQUARE 1/8
MILLER DISK
RETICULOCYTE COUNT
MILLER DISK
- SQUARE ISWHERE RETICS ARE COUNTED
LARGE SQUARE
RETICULOCYTE COUNT
MILLER DISK
- SQUARE IS WHERE RBCS ARE COUNTED
SMALL SQUARE 1/9
RETICULOCYTE COUNT
MILLER DISK
RBCS
MINIMUM OF - CELLS SHOULD BE COUNTED IN SMALL SQUARE
112 CELLS
RETICULOCYTE COUNT
MILLER DISK
RBCS
MINIMUM OF 112 CELLS IS EQUIVALENT TO - RED CELLS IN LARGE SQUARE
1008
RETICULOCYTE COUNT
MILLER DISK
FORMULA
RETICS= NO. OF RETICCS IN SQUARE A OR LARGE SQUARE X 100/ NO. OF RBCS IN SQUARE B OR SMALL SQUARE X 9
- BLOOD LOSS
- HEMOLYTIC ANEMIA
- TREATMENT TO PERNICIOUS ANEMIA, FOLIC ACID DEFICIENCY OR IRON DEFICIENCY
INCREASED RETICULOCYTES
MANUAL WBC COUNT
NORMAL WBC COUNT
4.0-11.0
X10^9/L
MANUAL WBC COUNT
USUAL DILUTION
1:20
MANUAL WBC COUNT
WBC DILUTING FLUIDS SHOULD BE - TO SWELL AND LYSE RBCS
HYPOTONIC
MANUAL WBC COUNT
WBC DILUTING FLUIDS (4)
- 1% AMMONIUM OXALATE
- 1% HYDROCHLORIC ACID
- 2-3% ACETIC ACID
- TURKS DILUTING FLUID
MANUAL WBC COUNT
ANTICIPATED WBC: 0.1-0.3
RECOMMENDED DILUTION:-
TYPE OF PIPETTE: -
1:10, WBC
MANUAL WBC COUNT
ANTICIPATED WBC: 3.1-30.0
RECOMMENDED DILUTION:-
TYPE OF PIPETTE: -
1:20, WBC
MANUAL WBC COUNT
ANTICIPATED WBC: >30.0
RECOMMENDED DILUTION:-
TYPE OF PIPETTE: -
1:100, RBC
MANUAL WBC COUNT
ANTICIPATED WBC: >100.0
RECOMMENDED DILUTION:-
TYPE OF PIPETTE: -
1:200, RBC
MANUAL WBC COUNT
MAKE A 1:20 DILUTION BY PLACING - UL OF BLOOD IN 475 UL OF WBC DILUTING FLUID
25UL
MANUAL WBC COUNT
MAKE A 1:20 DILUTION BY PLACING 25 UL OF BLOOD IN -UL OF WBC DILUTING FLUID
475UL
MANUAL WBC COUNT
ALLOW DILUTION TO SIT FOR - TO ENSURE RBC HAVE LYSED
SOLUTION WILL TURN (-COLOR)
10MINS, CLEAR
MANUAL WBC COUNT
CHARGE SIDES OF HEMACYTOMETER AT A -DEGREE ANGLE
45
MANUAL WBC COUNT
AFTER CHARGING, PLACE IN CHAMBER FOR - TO SETTLE
10MINS
THE - ARE COUNTED AS WBCS BC THEY ARE INDISTINGUISHABLE WHEN SEEN IN HEMACYTOMETER
NRBCS
WBC SHOULD BE CORRECTED WHEN NRBCS ARE
ADULT: - NRBC PER 100 WBC
> 5
WBC SHOULD BE CORRECTED WHEN NRBCS ARE
NEWBORN: - NRBC PER 100 WBC
> 10
CORRECTED WBC COUNT
FORMULA
UWBCC X 100/ NRBCS + 100
WBC DIFFERENTIAL COUNT
FOR ROUTINE TESTING, - SMEAR IS THE MOST WIDELY USED
WEDGE SMEAR
WBC DIFFERENTIAL COUNT
SCANNING / COUNTING METHODS
WBCS ARE COUNTED IN CONSECUTIVE FIELDS AS THE BLOOD FILM IS MOBED FROM SIDE TO SIDE
CROSS SECTIONAL/ CRENELLATION
WBC DIFFERENTIAL COUNT
SCANNING / COUNTING METHODS
WBCS ARE COUNTED IN CONSECUTIVE FIELDS FROM TAIL TOWARD THE HEAD OF THE SMEAR
LONGITUDINAL
WBC DIFFERENTIAL COUNT
SCANNING / COUNTING METHODS
BEGGINING NEAR THE TAIL ON A HORIZONTAL EDGE
BATTLEMENT/TRACK/BACK AND FORTH SERPENTINE
WBC DIFFERENTIAL COUNT
SCANNING / COUNTING METHODS
MOST PREFERRED COUNTING METHOD
BATTLEMENT/TRACK/BACK AND FORTH SERPENTINE
WBC DIFFERENTIAL COUNT
- DIFFERENTIAL COUNT
IF PX WBC COUNT IS <1.0X10^9/L
MULTIPLY RESULTS BY 2
50
WBC DIFFERENTIAL COUNT
- DIFFERENTIAL COUNT
ROUTINELY USED
100
WBC DIFFERENTIAL COUNT
- DIFFERENTIAL COUNT
OVER 10% EOSINOPHILS
OVER 2% BASOPHILS
OVER 11% MONOCYTES
MORE LYMPO THAN NEUTRO EXCEPT IN CHILDREN
200
WBC DIFFERENTIAL COUNT
- DIFFERENTIAL COUNT
WHEN WBC COUNT IS HIGHER THAN 40X10^9/L
200
WBC DIFFERENTIAL COUNT
- DIFFERENTIAL COUNT
WHEN WBC COUNT IS >100X10^9/L
300-400
WBC DIFFERENTIAL COUNT
NEUTROPHILS
RELATIVE COUNT: -
50-70
1.7-7.5
WBC DIFFERENTIAL COUNT
LYMPHOCYTES
RELATIVE COUNT: -
18-42%
1.0-3.2
WBC DIFFERENTIAL COUNT
MONOCYTES
RELATIVE COUNT: -
2-11%
0.1-1.3
WBC DIFFERENTIAL COUNT
EOSINOPHILS
RELATIVE COUNT: -
1-3%
0-0.3
WBC DIFFERENTIAL COUNT
BASOPHILS
RELATIVE COUNT: -
0-2%
0-0.2
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
4000-7000 WBC/UL
2-5 WBC/HPF
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
7000-10000 WBC/UL
4-6 WBC/HPF
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
10000-13000 WBC/UL
6-10 WBC/HPF
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
13000-18000 WBC/UL
10-20 WBC/HPF
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
WBC PER FIELD X - IF USING HPO
2,000
WBC DIFFERENTIAL COUNT
TOTAL WHITE CELL COUNT ESTIMATION
WBC PER FIELD X - IF USING OIO
3,000
WBC DIFFERENTIAL COUNT
PRESENCE OF IMMATURE GRANULOCYTIC CELLS
LEUKEMIA, BACTERIAL INFX
SHIFT TO THE -
LEFT
LIST OF SOME CONDITIONS ASSOCIATED WITH SPECIFIC INCREASE OF WBC
-APPENDICITIS
-BACTERIAL INFX
-PANCREATITIS
NEUTROPHILIA
LIST OF SOME CONDITIONS ASSOCIATED WITH SPECIFIC INCREASE OF WBC
-VIRAL INFX
-INFECTIOUS MONONUCLEOSIS
LYMPHOCYTOSIS
LIST OF SOME CONDITIONS ASSOCIATED WITH SPECIFIC INCREASE OF WBC
-THYPOID
-RICKETSSIAL INFX
-GAUCHERS
MONOCYTOSIS
LIST OF SOME CONDITIONS ASSOCIATED WITH SPECIFIC INCREASE OF WBC
-ALLERGIES
-PARASITIC INFX
EOSINOPHILIA
LIST OF SOME CONDITIONS ASSOCIATED WITH SPECIFIC INCREASE OF WBC
-ALLERGIC REACTION
-TYPE 1 HYPERSENSITIVITY
BASOPHILIA
HEMOGLOBIN DETERMINATION
THE - METHOD FOR HGB DETERMINATION IS THE REFERENCE METHOD
CYANMETHEMOGLOBIN
TEST USED TO DIAGNOSE AND FOLLOW TREATMENT OF ANEMIA
VARIES WITH
1. AGE
2. SEX
HGB DETERMINATION
HEMOGLOBIN DETERMINATION
NORMAL VALUES:
WOMEN: -
12-15 g/dL
HEMOGLOBIN DETERMINATION
NORMAL VALUES:
MEN: -
13.5-18 g/dL
HGB IS HIGHER IN THE - AND LOWER IN THE -
HIGH- MORNING
LOW-EVENING
HEMOGLOBIN DETERMINATION
- HGB
STRENOUS MUSCULAR ACTIVITY
SMOKER
HIGH ALTITUDE
INCREASED
HEMOGLOBIN DETERMINATION
- HGB
ANEMIA
AFTER 50 YRS OF AGE
LYING DOWN
DEHYDRATION
DECREASED
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
USES HCL
ACID HEMATIN, DIRECT VISUAL
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
USES NaOH
ALKALI HEMATIN, DIRECT VISUAL
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
ACID HEMATIN
HGB TURNS TO - HEMATIN
BROWN COLORED
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
ACID HEMATIN
ACID HEMATIIN + DW MATCH COLOR
YELLOW-BROWN
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
MOST RELIABLE AND RECOMMENDED METHOD FOR HGB
CYANMETHEMOGLOBIN, INDIRECT/PHOTOELECTRIC
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN USES - REAGENT
DRABKIN
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
COLOR INTENSITY: -
540 NM
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
ALL CAN BE MEASURED EXCEPT FOR - HEMOGLOBIN
SULFHEMOGLOBIN
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
OVERANTICOAGULATION - RESULTS
DOES NOT EFFECT
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
TURBIDITY WILL CAUSE - RESULTS
FALSELY ELEVATED
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
converts Hemoglobin (w/ ferrous iron) to methemoglobin
POTASSIUM FERRICYANIDE
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
converts methemoglobin to cyanmethemoglobin
POTASSIUM CYANIDE
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
improves lysis of RBC and decrease turbidity
NON IONIC DETERGENT
HEMOGLOBIN DETERMINATION
COLORIMETRIC METHODS:
CYANMETHEMOGLOBIN
allows the solution to be read after 3 minutes
DIHYDROGEN POTASSIUM PHOSPHATE
HEMOGLOBIN DETERMINATION
SPECTROPHOTOMETER: 540NM
SET -% TRANSMITTANCE
100%
HEMOGLOBIN DETERMINATION
DILUTION
1:251
HEMOGLOBIN DETERMINATION
STANDARD HGB CURVE
HEMOGLOBIN VALUE (G/DL)
X-AXIS
ABSCISSA
HEMOGLOBIN DETERMINATION
STANDARD HGB CURVE
OPTICAL DENSITY (ABSORBANCE)
Y-AXIS
ORDINATE
HEMOGLOBIN DETERMINATION
TURBIDITY=FALSELY ELEVATED DUE TO
HIGHER ABSORBANCE
HEMOGLOBIN DETERMINATION
REMEDY:
Centrifuge then read the absorbance of supernatant
High WBC count (>20x109
/L)
High platelet count (>700 x109
/L)
HEMOGLOBIN DETERMINATION
REMEDY:
make a 1:1 dilution with distilled water then multiply result by 2
Presence of Hb S or Hb C
HEMOGLOBIN DETERMINATION
REMEDY:
Use patient blank (0.01ml patient plasma + 5ml Drabkin’s reagent)
Lipemic sample
HEMOGLOBIN DETERMINATION
REMEDY:
Increase alkalinity of the hemiglobincyanide reagent by
adding potassium carbonate then repeat determination.
Abnormal globulins
HEMOGLOBIN DETERMINATION
If the drop sinks, its SG - that of copper sulfate
EQUAL/EXCEEDS
HEMOGLOBIN DETERMINATION
If the drop rises, its SG - that of copper sulfate
LESS
HEMOGLOBIN DETERMINATION
SPECIFIC GRAVITY OF COPPER SULFATE
1.053
HEMOGLOBIN DETERMINATION
ACCEPTABLE DROP OF BLOOD WILL SINK WITHIN 15SEC IF HB CONC IS -
> 12.5 G/DL
HEMOGLOBIN DETERMINATION
-METHOD
VAN SLYKE OXYGEN CAPACITY METHOD
GASOMETRIC METHOD
HEMOGLOBIN DETERMINATION
-METHOD
KENNEDY’S , WONG’S
CHEMICAL METHOD for IRON CONTENT
HEMOGLOBIN DETERMINATION
1 G OF HGB= -OXYGEN
1.34 ML
HEMOGLOBIN DETERMINATION
1 G OF HGB= -IRON
3.47 MG
- IS ABOUT 3X OF HGB VALUE
HEMATOCRIT
APPLIES ONLY ON NORMOCYTIC NORMOCHROMIC RBCS
DISCREPANCY=
FIRST INDICATION OF ERROR
RULE OF THREE
RULE OF THREE
HCT
HGB X 3
RULE OF THREE
HGB
RBC X 3
HCT/PCV
REPORTED AS -
36% OR
0.36 L/L
HEMATOCRIT DETERMINATION
ADULT MALES:
CU: 40-54%
SI: 0.40-0.54 L/L
HEMATOCRIT DETERMINATION
ADULT FEMALES:
CU: 35-49%
SI: 0.35-0.49 L/L
HEMATOCRIT DETERMINATION
NEWBORN
CU: 53-65%
SI: 0.53 TO 0.65 L/L
HEMATOCRIT DETERMINATION
-HCT
ANEMIA
HEMODILUTION
DECREASED
HEMATOCRIT DETERMINATION
- HCT
POLYCYTHEMIA
HEMOCONCETRATION
INCREASED
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
METHOD: WINTROBE AND LANDSBERG
ANTICOAGULANT: -
DOUBLE OXALATE
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
METHOD: VAN ALLEN, HADEN, SANFORD
ANTICOAGULANT: -
SODIUM OXALATE
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
METHOD: BRAY
ANTICOAGULANT: -
HEPARIN
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
SPEED OF CENTRIFUGATION
2000-2300G FOR 30MINS
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
WINTROBE TUBE: RED USED FOR -
- AT TOP
ESR, 0 AT TOP
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
WINTROBE TUBE: WHITE USED FOR -
- AT TOP
HEMATOCRIT, 10 AT TOP
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
WINTROBE TUBE
LENGTH: -
11.5 CM/115MM
HEMATOCRIT DETERMINATION
DIRECT METHOD (MACROMETHOD)
WINTROBE TUBE
BORE: -
3.0 MM
HEMATOCRIT DETERMINATION
DIRECT METHOD : -
SPX WHOLE BLOOD USING K2 EDTA OR CAPILLARY BLOOD
ADAM
HEMATOCRIT DETERMINATION
DIRECT METHOD: ADAM
- THOUGHT TO CAUSE 2-3% DECREASE IN HCT
K3 EDTA
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
LENGTH: -
7-7.5CM
70-75MM
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
BORE
1-1.2MM IN DIAMETER
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
CAN HOLF UP TO - OF BLOOD
0.05ML
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
PLUG: -
4-6MM
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
TYPE
WITH - BAND= WITH ANTICOAGULANT HEPARIN
RED
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
TYPE
WITH - BAND= WITHOUT ANTICOAGULANT
BLUE
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
CENTRIFUGE FOR - AT RCF 10,000-15,000XG
5 MINUTES
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
CENTRIFUGE FOR 5 MINS AT RCF -
10,000-15,000 XG
HEMATOCRIT DETERMINATION
CAPILLARY TUBE/MICROHEMATOCRIT TUBE
AFTER CENTRIFUGATION HCT MUST BE READ WITHIN -
10MINS
LAYERS OF SPUN HEMATOCRIT
Normall, layer is barely visible
In the presence of lipidemia, layer
several mm thick
TOP LAYER
FATTY LAYER
LAYERS OF SPUN HEMATOCRIT
Normally pale yellow and fairly clear
Excessive hemolysis results in cherry red
color
Jaundice produces a deep yellow color
SECOND LAYER
PLASMA
LAYERS OF SPUN HEMATOCRIT
Normally less than 1 mm thick
Thick layer when white cell count exceeds
10,000 cu/mm
Packed platelets are found in upper part
layer
THIRD LAYER
BUFFY COAT
LAYERS OF SPUN HEMATOCRIT
Volume (packed) is read as hematocrit
FOURTH LAYER
PACKED RED CELL
ERRORS IN MICROHEMATOCRIT RESULTS
FALSE - HCT
HEMOCONCENTRATION
DEHYDRATION
INSUFFICIENT CENTRIFUGATION TIME
BUFFY COAT IS READ
ALLOWING TUBE TO STAND LONGER
FALSE INCREASE
HEMATOCRIT DETERMINATION
TRAPPED PLASMA MAY CAUSE HCT TO FALSELY INCREASE AS MUCH AS -%
1-3%
THESE ARE CALCULATED TO DETERMINE AVERAGE VOLUME AND HGB CONTENT AND CONCENTRATION OF RBC IN SAMPLE
RBC INDICES
SERVES AS QUALITY CONTROL CHECK
USED FOR INITIAL CLASSIFICATION OF ANEMIA
RBC INDICES
RBC/ WINTROBE INDICES
32-36 G/DL
NORMOCHROMIC
RBC/ WINTROBE INDICES
<32
HYPOCHROMIC
RBC/ WINTROBE INDICES
> 36
HYPERCHROMIC
RBC/ WINTROBE INDICES
AN MCHC BETWEEN 36-38 G/DL SHOULD BE CHECKED FOR -
SPHEROCYTES
RBC/ WINTROBE INDICES
MCHC >38 CAUSED BY -
COLD AGGLUTININ
RBC/ WINTROBE INDICES
MCHC >38= COLD AGGLUTININ
INCUBATE SPX AT
37C, 15MINS
RBC/ WINTROBE INDICES
MCV: <80
MCHC: <32
MICROCYTIC, HYPOCHROMIC
RBC/ WINTROBE INDICES
MCV: 80-100
MCHC: 32-36
NORMOCYTIC, NORMOCHROMIC
RBC/ WINTROBE INDICES
MCV: >100
MCHC: 32-36
MACROCYTIC, NORMOCHROMIC
