Blood Smear Examination
Blood Smear Examination
Blood Smear Examination
EXAMINATION
Making Blood smear
PROCEDURE
placing a drop of blood from mixed
sample on a clean glass slide.
Spreader slide using another clean glass
slide at 30-40 degree angle.
high HCT
small angle
low HCT
large angle
A: Blood film with jagged tail made from a spreader with achipped
end.
B: Film which is too thick
C: Film which is too long, too wide, uneven thickness and made on
a greasy slide.
D: A well-made blood film.
Notes:
1.
2.
3.
4.
Principle
The main components of a Romanowsky stain are:
A cationic or basic dye (methylene blue or its oxidation
products such as azure B), which binds to anionic sites and
gives a blue-grey color to nucleic acids (DNA or RNA),
nucleoproteins, granules of basophils and weakly to granules
of neutrophils
An anionic or acidic dye such as eosin Y or eosin B, which
binds to cationic sites on proteins and gives an orange-red
color to hemoglobin and eosinophil granules.
pH value of phosphate buffer is very important.
Eosinophilic granules
Blue nucleus
Basophilic granules
Stain for 1-5 min. Experience will indicate the optimum time.
Add an equal amount of buffer solution and mix the stain by
blowing an eddy in the fluid.
Leave the mixture on the slide for 10-15 min.
Wash off by running water directly to the centre of the slide
to prevent a residue of precipitated stain.
Stand slide on end, and let dry in air.
Staining procedure
Thin smear are air dried.
Flood the smear with stain.
Stain for 1-5 min. Experience will indicate the
optimum time.
Add an equal amount of buffer solution and mix
the stain by blowing an eddy in the fluid.
Leave the mixture on the slide for 10-15 min.
Wash off by running water directly to the centre of
the slide to prevent a residue of precipitated stain.
Stand slide on end, and let dry in air.
TOO ACIDIC
SUITABLE
TOO BASIC
Correction:
1)
2)
3)
3. insufficient washing
4. alkaline pH of stain components
Correction :
1) check pH
2) shorten stain time
3) prolong buffering time
PERFORMING A MANUAL
DIFFERENTIAL AND ASSESSING RBC
MORPHOLOGY
PRINCIPLE
White Blood Cells.
1. Check for even distribution and
estimate the number present (also,
look for any gross abnormalities
present on the smear).
2. Perform the differential count.
PRINCIPLE
Red Blood Cells, Examine for :
1. Size and shape.
2. Relative hemoglobin content.
3. Polychromatophilia.
4. Inclusions.
5. Rouleaux formation or agglutination
Platelets.
1. Estimate number present.
2. Examine for morphologic abnormalities.
PROCEDURES
Observations Under 10
1. Check to see if there are good counting
areas available free of ragged edges and cell
clumps.
2. Check the WBC distribution over the
smear.
3. Check that the slide is properly stained.
4. Check for the presence of large platelets,
platelet clumps, and fibrin strands.
PLATELETS
Then
Observing direction:
LEUKOCYTOSIS
Leukocytosis, a WBC above 10,000 is usually due to
an increase in one of the five types of white blood
cells and is given the name of the cell that shows the
primary increase.
1. Neutrophilic leukocytosis
= neutrophilia
2. Lymphocytic leukocytosis
= lymphocytosis
3. Eosinophilic leukocytosis
= eosinophilia
4.Monocytic leukocytosis
=monocytosis
5.Basophilic leukocytosis
= basophilia
STAB NEUTROPHIL
Diameter:12-16
Cytoplasm : pink
Granules: primary
secondary
dense chromatin
BAND NEUTROPHIL
SEGMENTED NEUTROPHIL
Diameter: 12-16
Cytoplasm : pink
Granules: primary
secondary
SEGMENTED NEUTROPHIL
1.NEUTROPHILS
Neutrophils are so named because they are not well
stained by either eosin, a red acidic stain, or by
methylene blue, a basic or alkaline stain.
Neutrophils are also known as "segs", "PMNs" or
"polys" (polymorphonuclear).
2.
Granulocytic leukemia.
Typhoid fever
2.
Brucellosis
3.
Segmented neutrophile
Band neutrophil
EOSINOPHIL
Diameter: 14-16
Nucleus: blue
dense chromatin
2 lobes like a pair of glass
EOSINOPHIL
2. Parasitic infection
3. Eosinophilic leukemia
BASOPHIL
Diameter: 14-16
Cytoplasm : pink
BASOPHIL
Basophils
The purpose of basophils is not completely understood.
LYMPHOCYTE
LYMPHOCYTE
4.LYMPHOCYTES
Lymphocytes are the primary components of the
body's immune system. They are the source of
serum immunoglobulins and of cellular immune
response.
Two types of lymphocytes:
1. B lymphocyte : Humoral immunity
MONOCYTE
Diameter: 14-20
Cytoplasm : grey blue
Granules: dust-like lilac
color granules
Nucleus: blue
large irregularly shaped
and folded
MONOCYTE
NOTES
1. Do not count cells that are disintegrating
smudge cells
Basket cells
smudge cells
Basket cells
2- Abnormal differentials
1. 200 Cell diff:
labor unit)
5. If any cell type is extremely elevated (such as bands, monos, or eos >
20) indicate that you are aware of the abnormality by circling or
checking on the card next to the results.
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