WBC Identification Peripheral / Blood Smear

GRANULOCYTE A GRANULOCYTES

Neutrophil -

Lymphocyte
-

Eosinophil -

monocyte

Basophil
-

NEUTROPHIL
LYMPHOCYTE
90 -70% 20 -90%

10 -14mm
cell size -

cell size -

SL : 7 -10mm (equal to RBC size )

Nucleus -

Multi lobed
-

(1- Globes) , purple in color -

LL :
10 -14mm (double to RBC size )
-
_

cytoplasm -

Slight blush Nucleus -

single , very big ,

Oval / round in
-

Granules -
Fine, sand like particles , red brown or
purplish in color
=
shape central
,
in position occyupying
,

Function -

Phagocytosis ,
mediate febrile response whole of the cell

* high # of lobes semi or older

cytoplasm
-

scanty sky , blue in color , less

low # OF lobes juvenile

-

or
* younger amount than nucleus

Granules -
Absent

BASOPHIL Function -

produce antibodies

1- %

cell size -10 -14mm MONOCYTE

Nucleus -

-
Bilobed, purple in color , arranges sin shape
-
O 2 -8% cell size : 10 -18mm

cytoplasm -

Basophilio Blue in appearance

, Nucleus -

single, kidney shape pale , in color, peripheral in position

- -
- -

Granules -

Large , coarse , purple / blue overlying

,
nucleus cytoplasm -

Pale blue , amount more than the nucleus

Function -

Mild Phagocytosis Allergic manifestations Granules -

Absent
,

Function Phagocytosis work as tissue macrophages

-

,
EOSINOPHIL

1- 4%

cell size : 10 -14µm

Nucleus :
Bilobed , spectacle shape purple,
in color
-
-
-

granules :
Large ,
coarse , brick red in color , do
-
not cover nucleus
- -
-

Mild
Function
phagocytosis limiting allergic manifestations
:
, ,

provide local mucosal immunity

 HOW TO MAKE A PERFECT BLOOD SMEAR WEDGE SMEAR -

most common preparation

1) Thick to thin PERFORM DIFF COUNTING

2.) Occupy 2/3 OF the slide 1.) Optimal assessment Area

3) should not touch the edges

-

Between the thick (heel) and the feathery edge

A
-

RBCs uniformly and singly distributed

Angle
-

S
speed Few touching / overlapping
-
-

A -

Amount of blood
-

with central pallor

P -
pressure * locate the area that gives optional assessment for the diff

types of WBCS

•
locate the heel or thick part of the smears

the
feathery eye
•
to know if right area -
examine the red blood

cells

good area RBC still contain central

pallor
-
•

S shape is still intact

Thick smear -
no diff count → red blood cells will

be piling up on top of each other

-
cause distortion on cells increase WBC
,

classification will be difficult

-
evaluation will not be accurate

WBC DIFFERENTIAL COUNTING

-

AKA :
differential leukocyte count, pheripheral differential Feathery edge -
not good idea to count WBO

holes in the film

White blood cell morphology NBC differential Diff count
-

, ,

FORMAL NAME White Blood Cell Count Differential RBO 100k Flat
large
-
- :
,

body to respond to and eliminate distorted

PURPOSE :
asses the ability of the -
-

infections foreign substances (allergens) -

do not have central pallor

-

identify various stages of leukemia 27 Battlement pattern

COUNT -

100 WBCS

EXAMINE -

Morphology

IDENTIFY -

WBC type

ASSOCIATE -

Values to condition
WBO differential counter -
counted S classified

computer interfaced keypad advanced

-

more

Neutrophil -
multi lobed
-

-
kill bacteria S Foreign debris

Neutrophil ia -
bacteria or
fungal infection Lymphocyte -

deep staining nucleus

Lymphocytosis viral infection

-

-
eccentric

Fight viruses , make antibodies

-

Eosinophil -

bilobed with distinct red granules

-

killing parasites , cancer cells , involved in

allergic responses

Basophil -

bilobed but has distinct dark blue or

purple

granules
=
0.67×13.6
-

allergic reaction like eosinophil

monocyte -

kidney shaped nucleus

-
clean up damaged cells

absolute differential =
Relative differential ✗ WBC count
EXAMINATION OF BLOOD
CELLS
PERIPHERAL BLOOD SMEARS
INTRODUCTION
The examination of the peripheral blood smear is an integral part
of the routine examination of blood films. Essential information can be
obtained from this examination.
LEARNING OBJECTIVES
1. Evaluate the morphology of the different blood cells
2. Correlate the result of the examination with common hematologic
conditions
3. Distinguish between normal cells and abnormal cells
PRINCIPLE
To accomplish a blood film examination, the
microscopist prepares a “wedge-prep” blood film on a glass
microscope slide, allows it to dry, and fixes and stains it using
Wright or Wright-Giemsa stain.
The microscopist examines the RBCs and platelets by
light microscopy for abnormalities of shape, diameter, color,
or inclusions using the 50x or 100x oil immersion lens to
generate 500x or 1000x magnification.
PRINCIPLE
The microscopist then visually estimates the WBC count
and platelet count for comparison with their respective
instrument counts and investigates discrepancies.
Next, the microscopist systematically reviews, identifies,
and tabulates 100 (or more) WBCs to determine their percent
distribution. This process is referred to as determining the
WBC differential (“diff”).
MATERIALS
• Stained peripheral blood smears
• Microscope
• Oil for the oil immersion objective
PROCEDURE
1. Obtain blood smears from different individuals. / Prepared blood
smears.
2. Locate the area in the smear where the blood is not too thick or not
too thin. The ideal area is where the cells are not overlapping but
are not very far from each other
3. Focus under the LPO, and then shift to HPO and OIO.
4. Examine the morphology of the RBCs under the OIO. Observe the
WBCs and platelets as well.
WEDGE TECHNIQUE
An optimally stained smear has the following
characteristics:
1. The red blood cells (RBCs) should be pink to salmon.
2. Nuclei are dark blue to purple.
3. Cytoplasmic granules of neutrophils are lavender to lilac.
4. Cytoplasmic granules of basophils are dark blue to black.
5. Cytoplasmic granules of eosinophils are red to orange.
6. The area between the cells should be colorless, clean, and free of
precipitated stain.
RESULTS OF THE TEST
RESULTS OF THE TEST