White Blood Cells Morphology

Muhammad Imran khan

Lecturer Pathology
NWIHS
 Introduction to White blood cells
White blood cells are one cellular component of blood and are found through
the body, especially in the lymphatic system, as well as in the blood.
White blood cells are a part of the immune system and have an important role
in protecting the body against infection from pathogens and potentially harmful
foreign substances.
White blood cells can move out of the blood and into the tissues through a
process called diapedesis to fight local sites of infection and prevent its spread.
White blood cells are produced in the bone marrow, except some lymphocytes,
which are produced by lymphatic tissue.
The two main groups of white blood cells are the granulocytes and
agranulocytes.
 Granulocytes

Granulocytes are one of two group classifications of white blood

cells. Granulocytes generally have nuclei in two or more segments
and have granules that take on a distinctive color when stained for
examination under the microscope.

Granulocytes include:
Neutrophil
Eosinophil
Basophil
 Agranulocytes
or
Non-granulocytes
Agranulocytes are one of two group classifications of white blood
cells. Agranulocytes have nuclei in one segment.

Agranulocytes include:
Lymphocytes
Monocytes
 White blood cells

There are three types of

granulocyte named
according to their
staining characteristics
in blood films. They are
neutrophils,eosinophils
and basophils.
Mononuclear cells are
divided into
lymphocytes and
monocytes.
 Neutrophils
•Round in shape
•10-12u diameter
• Nucleus with 3-5 lobes connected by
threads of chromatin
• Cytoplasm contains fine purple
granules
• Concerned with Phagocytosis of
bacteria
• Hypersegmented neutrophils
contain six or more lobes
•Life span: 6 hours–few days
(days in spleen and other tissue)
 Band form Neutrophils

There are smaller numbers of

cells of neutrophil lineage
with non-segmented nuclei.
They are referred to as
neutrophil band cells or
band forms. They are less
mature than segmented
neutrophils.
An increased number of
band cells is referred to as
a'left shift'.
 Eosinophil
•Round in shape
•10-12u diameter
• Nucleus with 2 lobes (Bilobe)
connected by threads of
chromatin
• Cytoplasm contains course
red granules
• Eosinophils have a variety of
functions but are especially
important in defense against
parasitic infections.
• Life span: 8–12 days
(circulate for 4–5 hours)
 Basophil
• Round in shape
• 8-10u diameter
• Nucleus small kidney shape
• Cytoplasm contains dark blue
granules which obscure the cell
nucleus
• Basophils appear in many specific
kinds of inflammatory reactions,
particularly those that cause allergic
symptoms
• Basophils play a role in both
parasitic infections and allergies.
• Life span: A few hours to a few
days
 Normal lymphocyte
•Small lymphocyte:
•Round in shape
• Small 7-10u diameter
•Large condensed nucleus, with a scanty
bluish cytoplasm
• Nucleus is excentric
• Nucleus may show small indentation

•Large lymphocyte:
•Round in shape
•10-12u diameter
•Cytoplasm much greater than small
lymphocyte
•Cytoplasm blue in color

•Life span: Years for memory cells,

weeks for all else.
 Types of Lymphocytes
There are three types of lymphocytes known as:
T cells
B cells
natural killer cells

T cells get their name because they are developed in the thymus gland. These cells are
distinguished from other lymphocytes by the specialized T-cell receptor molecule that is
located on the surface of the cell. This molecule is important in immunity because it
recognizes antigens and is able to bind to them.
B cells are an important part of the immune system. B cells get their name because they
mature in the bone marrow of humans, These cells are distinguished from other lymphocytes
by a protein on their surface known as the B-cell receptor. This protein is specialized to
recognize and attach to specific antigens.
Natural killer cells are lymphocytes that are known to be cytotoxic. This means
that they have the ability to kill other cells. These cells are an important part of the immune
system because they are able to recognize virally infected cells, as well as some types of
tumor cells, and kill them before they cause a great amount of harm.
 Normal monocyte
• Monocytes are the largest WBC
•15-20u diameter
• Large kidney shape excentric
nucleus
• The cytoplasm is abundant, sky
blue in colou
• Some have vacuoles in the
cytoplasm

• Monocytes that leave the

bloodstream and enter the
connective tissue spaces are called
macrophages.

•These are famous for their

amoeboid movement and act as
scavanger cells.

• Life span: Hours to days

WBC Different types
 Types of white blood cells
Neutrophils: They kill and digest bacteria and fungi. They are the
most numerous type of white blood cell and your first line of defense when
infection strikes.

Eosinophils: They attack and kill parasites, destroy cancer cells, and help
with allergic responses.

Basophils: These small cells appear to sound an alarm when infectious agents
invade your blood. They secrete chemicals such as histamine, a marker of
allergic disease, that help control the body's immune response.

Lymphocytes.:They create antibodies to defend against bacteria, viruses, and

other potentially harmful invaders.

Monocytes: They have a longer lifespan than many white blood cells and
help to break down bacteria.
 What WBC are present in normal
peripheral blood?
In normal peripheral blood, there are three basic
types of leukocytes (illustrated below).
1. 2. lymphocytes
granulocytes normal atypical (few)
neutrophils
mature
band
eosinophils basophils 3. monocytes
 What methods can be used for the
differential WBC count ?

The differential WBC count and blood cell

study
may be performed by one of several
methods, e.g.:
• A manual microscopic examination of a Wright’s
(or Wright’s Giemsa) stained peripheral blood
smear.
OR
• An automated multi-channel instrument.
Manual Differential WBC
Count
 What kind of blood smear is used ?

A peripheral blood film (made from a drop of

blood from an EDTA anticoagulated tube or
skin puncture) is stained with Wright (or
Wright-Giemsa) stain and microscopically
examined using scanning (10x), high-dry
(40x), and oil (100x) objectives.
 How are the WBC identified and
classified?
Typical nuclear and cytoplasmic morphologic
features provide a means by which WBC can be
identified and classified as to:

• cell line (i.e., granulocytes [neutrophils, eosino-

phils, or basophils], lymphocytes, or mono-
cytes)
• maturity (i.e., mature cell or specific immature
stage of development).
• abnormal morphology (i.e., nuclear or cyto-
plasmic alterations)
 How are the WBC differentiated and
enumerated?
At least 100 WBC are counted and a tabulation is
made as to the number of each leukocytic cell
type included in the count.

The 100 cell count provides the RELATIVE

number (or percent) of each white blood cell
type present in the peripheral blood.
 Reference Range (Relative %)
Reference ranges (values considered to be normal)
for differential WBC counts may vary among
laboratories, but are usually about:

segmented band eosinophils lymphocyte monocytes

neutrophils neutrophils 1-5% s 20- 1-6%
50-70% 0- 5% 40%
 What terminology is used to indicate an
increased or decreased number of a
specific white blood cell line?

Cell Line Increased: Decreased:

Neutrophils Neutrophilia Neutropenia
Lymphocyte Lymphocytosi Lymphocytopeni

s Monocytes s
Monocytosis a
Monocytopenia
Eosinophils Eosinopenia

Basophils Eosinophilia
Basophilia Basopenia
Total WBC Count
vs.
Differential WBC
Count
 How does the differential WBC count
differ from the total WBC count?

The total WBC count reflects the total number of

all leukocytes in circulation but does not differ-
entiate leukocytes as to their various cell lines
(e.g., neutrophils, lymphocytes), stage of matur-
ity, (e.g., band, metamyelocyte), or abnormalities
when present (e.g., toxic granulation, hyperseg-
mented nuclei).

A differential WBC count must be performed to

provide that information.
Total WBC count = total number of
leukocytes without
differentiation:
“Diff” - differentiates and enumerates
WBC according to cell type, e.g.:

neutrophils, mature
lymphocytes
monocytes
eosinophils
basophils
 “Diff” - provides frequency distribution
(relative %and absolute number/L, e.g.
total WBC = 10,000/L):
lymphocytes
neutrophils, mature eosinophils
28%
65% 6500/L 3% 300/L
2800/L

basophils
1% 100/
L
monocytes
3%
300/L
 “Diff” - provides frequency distribution (relative %
and absolute number e.g., total WBC=50,000/L):

neutrophil, mature blast

50% 25000/L 1% 500/L

neutrophil, bands eosinophil

15% 7500/L 5%
2500/L
metamyelocyte
12% 6000/L basophil
3% 1500/L
myelocyte
7% 3500/L lymphocyte
5% 2500/L
promyelocyte,
2% 1000/L monocyte
0%
0/L
Relative % vs. Absolute
Number
 What is meant by the relative % and
absolute number of WBC ?

Relative % is based on the differential count of

100 white blood cells and reflects the per cent
of each cell type present in circulation.

If the total number of white cells in circulating

blood is known and the relative per cent of each
white cell type is known, then the absolute
number of each cell type per L of blood can be
calculated.
 continued:

For example: Given a patient with a total WBC

count of 8,000/L and the differential WBC count
shown below (i.e., the number observed for
each cell type in the 100 white cell count):

Segmented neutrophils 60%

Band neutrophils 5%
Lymphocytes 30%
Monocytes 2%
Eosinophils 2%
Basophils 1%
 continued:

Then the absolute number of each cell type/L

can be calculated by multiplying the per cent of
each cell type by the total number of WBC/L.
Segmented neutrophils 60% x 8,000 = 4,800
Band neutrophils 5% x 8,000 = 400
Lymphocytes 30% x 8,000 = 2,400
Monocytes 2% x 8,000 = 160
Eosinophils 2% x 8,000 = 160
Basophils 1% x 8,000 = 80
Total = 100% = 8,000
 What are the relative reference ranges
for leukocytes?
Cell Type Reference Ranges (Relative %)

Neutrophils 40-72 0-5

(bands)
(PMN)
Eosinophils 0-6

Basophils 0-0.2

Lymphocytes 24- 45

Monocytes 0.4 - 10
 Do the relative values always indicate
which cell line is or ?

If the total WBC count is “normal” (i.e., within

the established reference range), the relative
values are a good reflection of the number of
each cell type present, including increases
or decreases.

However, if the total WBC count is abnormal

(i.e., increased or decreased), the relative
percentage must be converted to an absolute
number of each cell type present in order to
determine which cell line is involved.
 How is the relative % converted to an
absolute number ?

For each white blood cell line, multiply the

percentage of cells counted (i.e., during the 100
cell count) by the total white blood cell count
to obtain the absolute number for the cell line.

Total WBC/L x relative % = absolute no./L

For example:
 Given a patient whose total WBC is 8,000/L,

and the relative distribution of leukocytes on

the peripheral blood smear is as shown below:

segmented band eosinophils lymphocyte monocytes

neutrophils neutrophils 2% s 25% 3%
65% 5%

How are the absolute numbers determined?

 How are the absolute numbers calculated
(total WBC = 8,000/L)?
Cell Type Relative % Absolute No.
65 (8,000 x 5,200/L
0.65)

Neutrophil (mature segmented)

5 (8,000 x 0.05) 400/L

Neutrophil (band)
 continued:
Cell Type Relative % Absolute No.

2 (8,000 x 0.02) 160/L

Eosinophil

25 (8,000 x 0.25) 2,000/L

Lymphocyte

3 (8,000 x 0.03) 240/L

Monocyte
 Total WBC 8,000/L x Relative %:
Cell Type Relative % Absolute No.
Neutrophil 65 5,200/L
(mature segmented)
Neutrophil (band) 5 400/L

Eosinophil 2 160/L

Lymphocyte 25 2,000/L

Monocyt e 3 240/L

Total 100 8,000/L

 What are the absolute reference ranges
for leukocytes?
The total WBC count reference range for adults is 4,500-
11,000/L at Charity & University Hospitals, but may vary
slightly among laboratories:
Reference Ranges
CELL TYPE RELATIVE % ABSOLUTE NO./L
Segmented neutrophils 42 - 72 1800 - 8000
Neutrophilic bands 0-5 0 - 550
Eosinophils 1-6 45 - 550
Basophils 0 – 0.2 0 - 200
Lymphocytes 24 - 45 1100 - 5000
Monocytes 0.4 - 10 200 - 1100
 However, given a patient whose total
WBC is 15,000/L,
with a relative distribution of leukocytes
on the peripheral blood smear the
same as the previous patient:

segmented band eosinophils lymphocytes monocytes

neutrophils neutrophils 2% 25% 3%
65% 5%

Is the interpretation the same for both patients?

 What are the calculated absolute
numbers in this case (i.e., total
WBC = 15,000/L)?
15000 x 0.65 = 9750 segmented neutrophils
15000 x 0.05 = 750 band neutrophils
15000 x 0.02 = 300 eosinophils
15000 x 0.25 = 3750 lymphocytes
15000 x 0.03 = 450 monocytes

The relative per cents are normal, but how do

these absolute numbers compare with the
established reference ranges?
 Are the absolute numbers still within
the reference ranges?
Absolute No./L
Cell Type Patient Reference Ranges
Neutrophils (mature) 9750 H 1800 - 8000
N Eutrophils (bands) 750 H 0 - 550
Eoinophils 300 0 - 600

3750 1100 - 5000

Lymphocyte
Monocytes 450 200 - 1100
 Comparison of relative %
and absolute numbers:
Cell Type Relative % Absolute No.
Neutrophil 65 N H 9,750/L
(mature segmented)
Neutrophil (band) 5 N H 750/L

Eosinophil 2 300/L

Lymphocyt 25 3,750/L
e Monocyte 3 450/L

Total 100 15,000/L

 Do the relative and absolute values have
the same interpretation in this case?

The relative percentages for all cell types for this patient
were within the reference range (i.e., normal).

However, in this case, there is neutrophila when

converted to absolute numbers based on a total WBC
count of 15,000/L because:

 segmented (mature) neutrophils = 9,750/L

which exceeds the reference range (1800 -
8000/L)
 band neutrophils = 750/L which also exceeds the
reference range (0 - 550/L).
THANKS