tlc count how different the cells

1. Dr Sandeep singh

2.  Smear prepration.
 Types of WBC and their counts
 Applied aspects.
 To draw a peripheral blood smear, stain it
with Leishman's stain and do the
differential count( DLC) of the given
blood sample.

3.  Crucial in the body defense against pathogens
Capable of phagocytosis.
 These are complete cells, with a nucleus and
organelles
 They lack Hb so they are colorless (i.e. white)
 Able to move into and out of blood vessels
• Can respond to chemicals released by
damaged tissues.

5. •Granules in their cytoplasm can be stained
inflammatory and allergic reactions.
• Neutrophils, Eosinophil, and Basophils

6. Agranulocytes
Lack visible cytoplasmic granules
Lymphocytes and Monocyte

7.  Normal levels =4,000 to 11,000 cells/mm3
Abnormal leukocyte levels
 Leucocytosis -Above 11,000 leukocytes/ml
Generally indicates an infection / Leukemia
• Leucopenia - low leukocyte level
Commonly caused by certain drug

13. 
Differential leukocyte count is performed on stained
blood film.
 Material required
 Glass slides
 Blood sample
 Leishman's stain
 Buffer water (pH = 6.8)
 Cedar wood oil
 Microscope with oil immersion objective

14.  Place a small drop of blood on one end of a clean slide, in
the middle. Hold a second slide (i.e., spreader) above the
first, right angle of 30 to 450.
 Move the spreader forward rapidly, thus spreading the
blood over the slide in a thin film.
 Leave the slide to air dry. A properly made film should dry
quickly; if the film is not dried rapidly there will be
rouleaux formation of RBC and the cells will appear
shrunken.
 A good blood film should not cover the entire surface of the
slide. It should be even, smooth and should be having three
parts i.e., head, body and tail.
 Dried film should be stained with Leishman's stain.

15.  Pour the Leishman's stain over the film to cover it completely. Keep it for two
minutes.
 After one minute, buffer water is added gently to the stain already on the blood
film to cover the slide completely (it is approximately double the volume of
stain).
 Allow the diluted stain to act for about 8 to 10 minutes.
 Wash the film in buffer water or distilled water. The film should be rose pink in
colour.
 Allow the film dry and focus under low power of microscope. Then put a drop
of cedar wood oil and see under oil immersion objective.
 Count the WBCs in a zig-zag fashion across the breadth of film, in the body
and tail junction area of film, until 100 leukocytes are counted.
 To record them draw a large square and divide it into 100 small squares.
Identify one cell and write in each square. Count different WBCs coming
across. The result thus obtained is directly in percentage.

19. N
L
RESULT:
Polymorph % =
Lymphocyte % =
Eosinophil % =
Monocyte % =
Basophil % =