Download MBBS (Bachelor of Medicine and Bachelor of Surgery) Practicals Latest Physiology Differential Leukocyte Count
Aim
Principle
Apparatus
Procedure
Precaution
PRINCIPLE:
BLOOD SMEAR IS PREPARED, STAINED WITH
LEISHMAN'S STAIN AND CELLS ARE IDENTIFIEDUNDER
OIL IMMERSION LENS.
4-5 GLASS SLIDES
LANCET/ SPRIT/COTTON
LEISHMAN'S STAIN
MICROSCOPE
CEDAR WOOD OIL
DISTILLED WATER
STAINING TRAY
preparation of blood smear
staining of blood smear
Examination of smear under oil immersion(100x) lens
selection of spreader
which has smooth edge, without coarse or uneven edge should
be avoided.
wash out the grease or oil, if it present in the glass slides as
well as at the edge of the spreader slide.
we have to use oil and grease free slides.
tongue shaped ( head ,body, tail)
should cover 2/3rd of the slide
Should not be thick
(single cell thickness)
should not have marks or
blank spaces in the smear.
Leishman's stain:
Belongs to Romanowsky group of stain.
Contains acidic and basic dye.
Composition:
Methylene blue- basic dye, positively charged and stains negatively charged
[acidic] particles ( stains nucleus of WBCs, the cytoplasm and basophilic
granules)
Eosin ?acidic dye, negatively charged and stains positively charged (basic)
particles (stains eosinophilic granules / RBCs)
Acetone free methyl alcohol-(fixative fix the smear to the slide)
WRIGHT STAIN
FIELD STAIN
MAKE SURE THE SLIDE IS DRY
POUR THE LEISHMAN'S STAIN DROP BY DROP
TILL IT COVERS ENTIRE SMEAR (8-10 DROPS)
NOTE THE TIME ALLOW FOR 1-2 MINs
(its known as FIXATION TIME)
ADD DOUBLE THE AMOUNT DROPS OF
DISTILLED WATER
WAIT FOR 6-8 MINS (FORMATION OF CATIONS
AND ANIONS OF BASIC AND ACIDIC DYE
REPECTIVELY) its knows as STAINING TIME.
TO DETERMINE THE DLC- (GRANULOCYTES
/AGRANULOCYTES)
TO STUDY THE MORPHOLOGY OF RBCS
DETECT THE PRESENCE OF PARASITES LIKE MALARIA,
FILARIA
SEX DETERMINATION CAN BE DONE BY IDENTIFICATION
OF BAR BODY
FOCUS UNDER HIGH POWER (100X)
PUT ONE DROP OF CEDAR WOOD OIL
1 0 - 1 4
CYTOPLASM PINK IN COLOUR
NUCLEUS HAVING 2 - 5 LOBES
HAVING FINE PINK OR PURPLE GRANULES
PURPULE/PINK IN COLOUR
1 0 - 1 4 MICRON/ NUCLEUS BILOBED PURPULE
COARSE ORANGE TO RED IN COLOUR
BLUE COLOUR
GRANULES/ CYTOPLASM PINK COLOUR
18-22/NOTCHED OR OVAL OR HORSE SHOE SHAPE
8 - 1 0 /NUCLEUS 2 - 3 LOBES/NOTPROPERLY
NUCLEUS
VISIBLE BECAUSE OF GRANULES/
NONE OR MODERATE COARSE AZURE GRANULES
COARSE BLUISH BLACK GRANULES OVERLYING
THE NUCLEUS
CYTOPLASM SKY BLUE
LARGE LYMPHOCYTE
SMALL LYMPHOCYTE
1 0 - 1 4
7 - 1 0
SMALL ROUND NUCLEUS
NUCLEUS LARGE ROUND FILLS THE WHOLE CELL
CYTOPLASM SKY BLUE
SKY BLUE THIN RIM
GRANEULS NONE OR FEW
SKY BLUE/NONE OR MODERATEGRANEULES
CELLS
SIZE
NUCLEUS
CYTOPLASM
GRANULES
NEUTROPHIL
1 0 - 1 4
2 - 5 LOBES
PINK
FINE PINK OR PURPLE
PURPULE/PIN
GRANULES
K
EOSINOPHIL
1 0 - 1 4 MICRON
BILOBED PURPULE
PINK
COARSE RED IN
BLUE COLOUR
COLOUR
BASOPHIL
8 - 1 0
2 - 3 LOBES/NOT
PALE PINK
COARSE BLUISH BLACK
PROPERLY VISIBLE
GRANULES OVERLYING
BECAUSE OF
THE NUCLEUS
GRANULES
LARGE LYMPHOCYTE
1 0 - 1 4
SMALL ROUND
SKY BLUE
NONE OR FEW
NUCLEUS
SMALL LYMPHOCYTE
7 - 1 0
LARGE ROUND FILLS
SKY BLUE THIN RIM
SKY BLUE/NONE OR
THE WHOLE CELL
MODERATE
GRANEULES
MONOCYTE
1 8 - 2 2
NOTCHED OR OVAL
SKY BLUE
NONE OR MODERATE
OR HORSE SHOE
COARSE AZURE
SHAPE
GRANULES
NEUTROPHIL
50-70%
LYPHOCYTE
20-40%
MONOCYTE
2-8%
EOSINOPHIL
1-4%
BASOPHIL
0-1%
DRAW 100 SQUARES
IDENTIFY THE VARIOUS CELLS ENTER FIRST LETTER
N-NEUTROHIL
E-EOSINOPHIL
B-BASOPHIL
L- LYMPHOCYTE
M-MONOCYTE
NEUTROPHIL=
? %
EOSINOPHIL =
? %
LYMPHOCYTE=
? %
MONOCYTE =
? %
BASOPHILS =
? %
ABSOLUTE NEUTROPHIL =WBC count x neutrophil %= - - c u m m
100
DETERMINATION OF THE PERCENTAGE DISTRIBUTION OF DIFFERENT
TYPES OF NEUTROPHILS ON THE BASIS OF NUMBER LOBES IN THEIR
NUCLEUS
Normal count
N1 - 2-10%
N2 - 20-30%
N3 - 40-50%
N4 ? 10-15%
N5 ? 2-5 %
SHIFT TO LEFT ?MORE YOUNGER CELLS (REGENARATIVE
SHIFT)
SHIFT TO RIGHT - MORE OLDER CELLS (DEGENARATIVE
SHIFT)
Textbook of Practical Physiology by
A.K.Jain
C.L. Ghai &
G.K.Pal
Net Source for images
This post was last modified on 30 November 2021