White Blood Cells Count (WBC) & Differencial Count (DC) : Liu, Liu

WHITE BLOOD CELLS COUNT
(WBC)
&
DIFFERENCIAL COUNT (DC)
Liu, liu
First Teaching Hospital
ZhengZhou Univ.
1
Liu,
Liu,Yan-Fang
Yan-FangMD,
MD,Ph.
Ph.D.D.First
FirstTeaching
TeachingHospital
HospitalZhengZhou
ZhengZhouUniv
Univ
W B C – Normal value
Adult: 4.0 ～ 10.0×109/L;
New born baby: 15.0 ～ 20.0 × 109/L;
Infant (6m ～ 2ys): 11.0 ～ 12.0 ×

109/L.
2
Liu, Yan-Fang MD, Ph. D. First Teaching Hospital ZhengZhou Univ
W B C – Clinical significance
• Leucocytosis : WBC > 10.0 × 109/L;
• Leucopenia : WBC < 4.0 × 109/L.
The increase or decrease of total numbers o

f WBC is mainly influenced by neutrophili
c granulocytes.
3
Differential count -- Methods
• 1. Making Blood smear:
Take a drop of blood, put on a slide, place anothor slide
over the blood and smear across the first slide .
• 2. Staining:
After the smear dry, flood the Wright’s, fixed for 1 ～ 2
min. Alter the relative amounts of buffer, stain for 5 ～
10min, and wash the smear with water, to remove the st
ains. Air dry.
• 3.Examination:
Directly under oil immersion. Record the different kind
s WBC till the total number reaching 100.
4
Classification of WBC:
• Granulocytes
• Neutrophil,
• Eosinophil,
• Basophil,
• Lymphocytes,
• Monocytes.
5
Differential count -- Normal value
• Neutrophil:
• Segmental 0.50 ～ 0.70 (50 ～ 70%);
• Stab 0.01 ～ 0.05 (1 ～ 5%);
• Eosinophil: 0.005 ～ 0.05 (0.5 ～ 5%);
• Basophil: 0 ～ 0.001 (0 ～ 1%);
• Lymphocyte: 0.20 ～ 0.40 (20 ～ 40%);
• Monocyte: 0.03 ～ 0.08 (3 ～ 8%).
6
Development of neutrophils
7
Neutrophils
• Occurance
– 60-70% of all white blood cells
– Most common leukocyte
• Appearance
– Multilobed nucleus
• 3-5 lobes
– “clear” granules
– 10-15 m
8
Neutrophilic stab granulocyte
9
Neutrophilic segmental granulocyte
10
Neutrophil function
• Earliest to arrive in an infection
• Main component of pus
• Professional phagocyte
– Opsonization increases phagocytic activity
• Coating with antibodie receptors and complement receptors
• Highly motile cells
– Respond to chemotactic factors
• released from damaged tissue
– Leave blood circulation
11
Increases ( neutrophilia)
-- Physiological
Fetus and neonatal stage (10.0 ～ 20.0)
Pregnant women above 5m(>15.0);
In late Pregnancy may reach 20.0;
Heavy manual labor or violent exercise;

exposure to the cold air for long Time (rain,
snow,wine)
Heavy meal or shower bath.

12
Increases ( neutrophilia)
-- Pathological:
(1) Acute infection: especially the pyogenic
staphylococcus aurous, streptococcus hemolyticus, diploco
ccus pneumonia >20.0.
(2) Severe tissue injury
Major operation ; acute myocardial infraction.
(3) Acute massive hemorrhage and destruction of
a large amount of RBC.
(4) Acute poison
Disturbance of metabolism, acute chemical poisoning
.
(5) After transplantation of organs.
(6) Leukemia and malignant tumor.
13
Decreases (neutropenia) - 1
(1)Certain infections
Some Gram(-) bacilli infection :
Typhoid fever ,Paratyphoid fever
Virus infection:Influenza
Protozoa infection:
Malaria
Kala-azar
14
Decreases (neutropenia ) - 2
(2)Certain blood diseases:

Aplastic anemia, some kind of leukemia
Acute granulocyte deficiency syndrome.
(2)Chronic physical and chemical injury:
Long term contact lead, Mercury, Benzene.Long ter
m exposure to x-ray, after radiotherapy
Administration of BM suppressing drug &
anti-cancer drug.
.
15
Decreases (neutropenia ) - 3
4). Auto-immunological diseases :

Systemic Lupus Erythematosus (SLE) .
(5). Hyper-function of spleen :
Splenomegaly of various causes may develop le
ucopenia such as portal cirrhosis of liver , Bant
i’s syndrome.
16
Eosinophils
• Occurance
• 0.5~5% of all white blood cells
• Appearance
– “red-orange” granules
• 10-15 m
• Function
– Phagocytic cells with an affinity for parasites
– Receptors for IgE
• Plays a role in decreasing hypersensitivity reactions
17
Eosinophil(segmental)
18
Eosinophilia Eosinopenia
Allergic reaction • Acute stress
Parasites • Certion stage of typhoi
dermatosis d
Some kinds of carcinoma • Steroids/Cushing’s
Some hematologic malig syndrome
nant desease
others
19
Basophils
Occurance
• <1% of all white blood cells
• Least common leukocyte
Appearance
– Big “blue” granules
– 10-15 m
Function
– Immediate hypersensitivity responses (allergies)
• Histamine and other mediators of inflammatory
response in granules
20
Basophil(segmental)
21
Basophilia Basopenia
• Hypersensitivity
reactions • No significance
- asthma, eczema
• Hematologic
malignancy
• Ulcerative colitis. pox
22
Monocytes
• Occurance
– 3~8% of WBC
• Appearance
– Large nucleus
• Horseshoe or kidneybean shape
• About 50% of the cell
– small granules
– 10-20 m
23
monocyte
24
Monocyte function
• Tranform into macrophages
– After leaving bloodstream
– More phagocytic than neutrophils
– Primary antigen processing and presenting cell
– Responsible for expressing MHC class II to CD4
T cells(major histocompability complex )
• Monocyte-macrophage system
– Skin--Langerhan’s cell
– Bone--Osteoclast
– Liver--Kuppfer cell
– Brain--Microglia
25
Monocytosis Monocytopenia
Physiological:
child,infant • No significance
pathological
• Infection: subacute bact
erial endocarditis, tuber
culosis
• Hematological desease
-acute leukemia,MDS
26
Lymphocytes
• Occurance
– 20~40% of all white blood cells
• Appearance
– Round nucleus
• about 90% of cell
– 8-10 m
– Cannot distinguish between B and T lymphocytes
under microscope
27
Macrolymphocyte and small lympho
cyte
28
Lymphocyte function
– B lymphocytes
• become plasma cells
– produce antibodies (humoral immunity)
• B cells formed in bone marrow and travel to
lymph nodes and spleen
– T cells
• migrate from bone marrow and mature in
thymus
• Have many immune functions(cell immunity)
29
Lymphocytosis Lymphopenia
• Count varies with age • Decreased production
• Viral infection – Inherited immune defic
• Other infections iency.
– Tuberculosis,whooping co – AIDS(acquired)
ugh • Increased destruction
• Hemotological desease: a – Steroids/ Cushing’s
cute leukemia; – Radiation, some drugs
lymphoma
others:carcinoma,
graft rejection
30
The change of nuclear picture o
f neutrophil granulocyte
Nucleus shifting to the left:
Increase of stab cell with the appearance o
f metamyelocyte, myelocyte and promyelo-cyt
e, etc.
The total numbers of WBC may be increased

(frequently)/ normal / reduced (such as typhoid
fever, virus infection, malaria).
31
THE SEVERITY OF SHIFT TO LEFT
• Slight shifting to the left:
Stab cells > 6%, no immature-myelocyte; common infe
ction, hemorrhage, hemolytic diseases.
• Middle shifting to the left:
Stab cells > 10%, a few immature-myelocyte; severe i
nfection, acute hemolytic, poisoning.
• Severe shifting to the left:
Stab cells > 25%, more immature-myelocyte; myelocyti
c leukemia.
32
Change of nucleus
33
Nucleus shifting to the left
34
Nucleus shifting to the right
5 lobed nucleus granulocyte beyond 3%
Always Accompanied by decrease of WBC mani

festation deterioration of hemopoiesis function.
Also can be seen in megaloblast ane
mia,use of some antimetabolism dru
gs,convalescent period of inflammat
ion
35
Nucleus shifting to the right
36
Common pathological of WBC
toxic appearance – 1
(1). Size is not uniform:
Large /small is not average.
Cause: Long term pyogenic inflammation or

chronic infection may leads the irregular div
ision of immature cells, or the effect of chem
otatic factors, endotoxin, etc.
37
Size is not uniform
38
toxic appearance-2
(2). Toxic granules:
Blue black granules appearance in the cytoplasm (is a
n interference in process of formation of special gran
ule or deterioration of the granules).
(3). Vacuole retrogression:
Single / several vacuoles appeared in the cytoplasm (f
requently seen in severe infection, especially septicem
ia).
Cause: Injury of cells leads to the fatty degeneration

of cytoplasm.
39
Toxic granules
40
Vacuole retrogression
41
toxic appearance - 3
(4). Nucleus retrogression:
Pyknotic change -- Appears as a homogeneous
dark purple lump;
Lysis of nucleus -- Swells and stains lightly freq
uently accompanied by break of nuclear membrane;
Break of nucleus -- Nuclear breaks to several pi
eces.
Clinical significance is same as above.

42
Nucleus retrogression
43
Staff body ( Aure’s body)
Appears as a red slender staff like substance

in cytoplasm, one or several in number and
1-6  in length.
It is only seen in AML.
44
Auer bodies
45
Atypical form of lymphocyte - 1
• Abnormal morphology of lymphocytes may be
seen occasionally in a blood smear of normal
person(<2%), this kind cell can be classified
into 3 types.
• Owing to the progress of immunology, atypical

cells are known as “T” lymphocytes. It is a
result of stimulating of the virus or allergen
/hypersensitive antigen.
46
• Type 1 (vacuole type):

Cell body larger than normal lympho
cyte,
Cytoplasm deep blue with numerous s
mall vacuoles and minute azurophil gran
ules.
47
• Type 2 (irregular type ):

Cell body is as large as monocyte,
Nucleus is round or irregular, the chrom
atin is not clear,
Cytoplasm is abundance, light blue / b
lue in color, no vacuole , small amount az
urophil granules.
48
• Type 3 (juvenile type ):

Cell body is larger,
1-2 nucleoli may be seen ,
Cytoplasm is deep blue and with a few
vacuoles.
49
Atypical lymphocyte-vacuole type
50
Atypical lymphocyte-irregular type
51
Atypical lymphocyte-juvenile type
52
Clinical significance:
• >10% is significant in differential diagno
sis of infectious mononucleosis.
• It is may be seen in virus infection, such

as pediatric rubella, pneumonia; and bac
terial infection, rickettsia （立克次体病 )
, or hypersensitive diseases ( >2%).
53

White Blood Cells Count (WBC) & Differencial Count (DC) : Liu, Liu

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WHITE BLOOD CELLS COUNT

Adult: 4.0 ～ 10.0×109/L;

New born baby: 15.0 ～ 20.0 × 109/L;

Infant (6m ～ 2ys): 11.0 ～ 12.0 ×

• Leucocytosis : WBC > 10.0 × 109/L;

• Leucopenia : WBC < 4.0 × 109/L.

The increase or decrease of total numbers o

Heavy manual labor or violent exercise;

Heavy meal or shower bath.

(2)Certain blood diseases:

4). Auto-immunological diseases :

The total numbers of WBC may be increased

Always Accompanied by decrease of WBC mani

Cause: Long term pyogenic inflammation or

Cause: Injury of cells leads to the fatty degeneration

Clinical significance is same as above.

Appears as a red slender staff like substance

It is only seen in AML.

• Owing to the progress of immunology, atypical

• Type 1 (vacuole type):

• Type 2 (irregular type ):

• Type 3 (juvenile type ):

• It is may be seen in virus infection, such

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