Leukocyte Total and

Differential Count
Lab 5
Classification
 Granulocytes:
1- Neutrophils:
A- Band
B- Segment
2- Eosinophils
3- Basophils
 Mononuclear:
1- Lymphocytes
2- Monocytes
Band
 Nucleus: Indentation is >50% of the width
of the nucleus with abundant granules.
Segment
 Nucleus: 2-5 lobes connected by thin
filaments without visible chromatin.
 Cytoplasm: Pale pink, cream colored or
colorless with abundant fine neutral
granules.
 Eosinophil
 Size: 12-17 um.
 Nucleus: 2 – 5 lobes (the majority of mature cells
have 2 – 3 lobes) connected by thin filaments without
visible chromatin with abundant pale orange to deep
orange granules.
 Basophil
 Nucleus: Usually 2 lobes connected by thin filaments without
visible chromatin. With granules variable in number with uneven
distribution, may obscure the nucleus; deep purple to black
Lymphocyte
 Nucleus: Round to oval; may be slightly indented.
 Cytoplasm: Scant to moderate; sky blue
 Granules: None in small lymphocytes may be a few
azurophilic in larger lymphocytes; if granules are
prominent, the cell is called a large granular
lymphocytes.
Monocyte
 Nucleus: Variable; may be round, horseshoe
shaped, or kidney shaped; often has folds
producing brain-like convolutions.
 Granules: Many fine granules frequently giving
the appearance of ground glass.
 Vacuoles: Absent to numerous.
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11
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Normal WBCs some examples

(1) (2)

(4) 13
14
15
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 REPORTING A DIFFERENTIAL COUNT

By relative % of each WBC type

(INCORRECT)
Ways to
report the
values

By absolute number of each WBC type

(CORRECT)
(%of each cell type x total WBC number)

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Differential WBCs
Neutrophils 2.0 – 7.0 40 – 80 %

Lymphocytes 1.5 – 4.0 20 – 40 %

Monocytes 0.2 – 1.0 2 – 10 %

Eosinophils 0.02 – 0.5 1–6%

Basophils 0.02 – 0.1 0–1%

 EXAMPLE

• nº1 : 70% segmented and 10.0 x 109/L WBCs

• nº2: 70% segmented and 1.0 x 109/L WBCs
• nº3 : 70% segmented and 50.0 x 109/L WBCs
% Absolute segmented
Segmented neutrophils neutrophils x 109/L

• nº1 : 70 (NORMAL) 7.0 NORMAL

• nº2: 70 (NORMAL) 0.7 (DECREASED)

• nº3 : 70 (NORMAL) 35.0 (INCREASED)

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 INTERPRETING DIFFERENTIAL RESULTS

Neutrophilia
Eosinophilia
 in a particular leukocyte Basophilia
Lymphocytosis
Monocytosis

Suffix “penia”
 in a particular leukocyte (i.e. Lymphopenia)

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Left-shift and right-shift of
neutrophil:
 Left-shift: non-segmented neutrophil >
5% or at least one more immature cell
 Right-shift: hypersegmented neutrophil
Leukemoid reaction
 Definition: marked leukocytosis with shift to
left

 Toxic granulation – vaculation

 LAP : ↑↑
 No clonality (immunophenotyping and ph)
Leukemia
 Cancer of the blood or bone
marrow characterized by an abnormal
increase of immature white blood
cells called "blasts“ which replace the
normal hemopoietic cells.
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Classification
Cell type Acute Chronic

Lymphocytic leukemia Acute lymphoblastic Chronic lymphocytic

(or "lymphoblastic") leukemia (ALL) leukemia (CLL)

Myelogenous leukemia Acute myelogenous

Chronic myelogenous
(also "myeloid" or leukemia (AML)
leukemia (CML)
"nonlymphocytic") (or myeloblastic)
Acute leukemia
 rapid increase in the number of immature
blood cells
 Predisposing factors of acute
leukaemia
 1-Radiation .
 2-Chemical e.g cytotoxic drugs.
 3-Viruses .
 4-Genetic factor and chromosomal
abnormalities e.g Down syndrome,
Fanconi anaemia.
 5-Immunological factors e.g
immunodeficiency states.
 6-Predisposing haematological disease
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Clinical Picture
 1-Due to bone marrow failure
 Anaemia:pallor,lethargy,dyspnea.
 Infection :fever,malaise,sign of mouth,
throat,skin and respiratory infection.
 Thrombocytopenia:bleeding,spontaneous
bruises,purpura,bleeding gums,
menorrhagia and internal haemorrhage.

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Clinical Picture
 2-Due to organ infiltration
 Tender bones(sternum) especially in
children
 Lymphadenopathy in ALL
 Moderate splenomegaly and
hepatomegaly specially in ALL
 Gum infiltration and skin lesion in M4 and
M5
 Renal failure and hypokalemia in M4 and
M5 due to release of lysozyme
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 Meningeal and other organ infiltration
Acute leukemia Diagnosis
 CBC
o WBCs: ↑, N or ↓
o Anemia and thrombocytopenia
 Blood film or BM: Blasts > 20%
 Cytochemistry: MPO –SBB – TdT
 Immunophenotyping
 Cytogenetics (chromosomal abnormalities)
 Electron microscopic (important for M0 and M7)
 AML
Type Name

M0 acute myeloblastic leukemia, minimally differentiated

M1 acute myeloblastic leukemia, without maturation

M2 acute myeloblastic leukemia, with granulocytic maturation

M3 promyelocytic, or acute promyelocytic leukemia (APL)

M4 acute myelomonocytic leukemia

acute monoblastic leukemia (M5a) or acute monocytic

M5
leukemia (M5b)

acute erythroid leukemias, including erythroleukemia (M6a)

M6
and very rare pure erythroid leukemia (M6b)

M7 acute megakaryoblastic leukemia

ALL
 ALL-L1: small uniform cells
 ALL-L2: large varied cells
 ALL-L3: large uniform cells
with vacuoles (bubble-like features)
Chronic leukemia
 Excessive relatively mature white blood
cells. Typically taking months or years to
progress, the cells are produced at a much
higher rate than normal,
Chronic leukemia
 CBC:
o Leukocytosis
o Anemia and thrombocytopenia: late and
mild
CML

 Blood film: Leukocytosis –

shift to left – eosinophilia –
basophilia
 LAP: ↓ or absent
 Philadelphia chromosome.
Philadelphia chromosome
 specific chromosomal abnormality that is
associated with chronic myelogenous
leukemia (CML). It is the result of a
reciprocal translocation between
chromosome 9 and 22
CLL
 Persistant lymphocytosis > 5000
 Mature lymphocytes
 Anemia and thrombocytopenia: late and
mild
 Immunophenotyping