Blood cell counting

1. BLOOD CELL COUNT
RAJESH MOHESS,CLT

2. BAND (Stab cell)
• Band are round or oval in shape and smaller than metamyelocyte.
• They can be found in low percentage in the peripheral blood smear 0% -
5%
• Bone Marrow: 17% - 33%
• Size: 9 to 15 μm
• Chromatin: The nucleus has a deeply indented U-shape, C, S, kidney-bean
shape or rod shape of equal thickness
• The chromatin is coarse and clumped and is stained dark red or red blue.
No nucleoli
• They are often described as ‘stab’ (rod-like) forms.
• Cytoplasm: Brown-pink (light acidophilic) and contains secondary
granules characteristic for some species of granulocytes
• Differentiating characteristics: No filament
It may resemble a metamyelocyte but indentation is more severe and
chromatin is more clumped

3. BAND (Stab cell)

4. MATURE NEUTROPHIL
(Segmented Neutrophil/PMN)
• Segmented neutrophilic granulocyte is a mature cell, round or
oval shape
• Size: 9 to 15 μm
• Chromatin: The nucleus is lobulated, with two to five lobes
connected by a thin chromatin thread-like filament
• The chromatin stains purple and is coarse and arranged in clumps.
• The nucleus of 1-16 % of neutrophils may have a drumstick-shaped
appendage attached to one lobe by a chromatin strand.
• Neucleoli are absent
• Cytoplasm is pale blue to pink granules
• N:C 3:1 in favor of the cytoplasm.
• Distinguishing characteristics: if no constriction is observed in
nucleus, then the cell is a band
• Bone Marrow: 3% - 11%
• Peripheral smear: 50 – 70 % (adults)

5. NEUTROPHIL

6. NEUTROPENIA
Neutropenia - low neutrophil count in the blood
Neutrophils serve as the major defense of the body
against aaccuuttee bbaacctteerriiaall and certain fungal infections.
 Neutrophils usually constitute about 50 to 70% of all
white blood cells in the bloodstream.
When the neutrophil count falls below 1,000 cells per
microliter of blood, the risk of infection increases
Without the key defense provided by neutrophils,
people have problems controlling infections and are at
risk of dying from an infection

7. CAUSES OF NEUTROPENIA
Neutropenia significantly increases the risk of life-threatening
infection
Neutropenia is often caused by cancer chemotherapy or radiation
therapy
Neutropenia can develop if neutrophils are used up or destroyed in
the bloodstream faster than the bone marrow can make new ones.
With some bacterial infections, some allergic disorders, and some
drug treatments, neutrophils are destroyed faster than they are
produced.
People with an autoimmune disease can make antibodies that
destroy neutrophils and result in neutropenia.
People with an enlarged spleen may have a low neutrophil count
because the enlarged spleen traps and destroys neutrophils.
Many drugs, including phenytoin, chloramphenicol, sulfa drugs,
and many drugs used in cancer treatment (chemotherapy)

8. NEUTROPHILIA
Neutrophilia (Neutrophilic Leukocytosis) is a higher than
normal Neutrophils in the blood
Infections by bacteria, viruses, fungi, and parasites may all
increase the number of neutrophils in the blood.
The number may rise in people who have an injury, such as
a hip fracture or burn.
Inflammatory disorders, including autoimmune disorders
such as rheumatoid arthritis, can cause an increase in the
number and activity of neutrophils.
Some drugs, such as corticosteroids, also lead to an
increased number of neutrophils in the blood.
Myelocytic leukemias can lead to an increased number of
immature or mature neutrophils in the blood

10. EOSINOPHIL
• Eosinophils are round or oval cells that are, on average,
slightly larger than the mature neutrophils.
• Size: 10 to 16 μm
• N:C: Barely 1:1
• Chromatin: Eccentric nucleus, usually bilobed
• Nucleoli is absent
• Cytoplasm is large and orange-pink with distinctive red-orange
specific granules which are highly metabolic and
contain histamine and other substances
• Distinguishing characteristics: Granules are uniformly round,
large, and individualized; if stain is less than adequate,
observe granules carefully for their crystalloid nature
• Bone Marrow: 0% - 3%
• Peripheral smear: 0% - 4% (adults)

11. EOSINOPHIL

12. EOSINOPHIL DISORDERS
• Eosinophilic granulocytes are mostly located in tissues, and
not in the peripheral blood.
• Before moving to tissues they remain in the peripheral blood
for about 10 hours.
• Eosinophilia accompanies immune, allergic reactions and
parasitic reactions.
• Eosinophilic granulocytes have antihistaminic function –
preventing edema and smooth muscles spasm.
• They have power of phagocytosis, but not as great as the
neutrophils
• Eosinopenia can occur with Cushing's syndrome, stress
reactions, and treatment with corticosteroids

14. BASOPHIL
• The mature basophilic is a round or oval cell.
• Size: 10 to 14 μm
• N:C: Difficult to determine
• Nucleus is round, kidney-shaped or uneven, with a larger number of
notches, and occupies the major part of the cell.
• The nucleus is segmented into different lobes connected by chromatin
filaments
• Chromatin: Coarse, clumped bilobed
• Nucleoli are absent
• Cytoplasm: Rose- or lavender-colored or sometimes does not stain.
• Has large, round, deeply stained blue to black unevenly distributed
granules that may overlap and obscure the nucleus.
• Distinguishing characteristics: Size and color of granules will obscure the
nucleus
• Bone Marrow: <1%
• Peripheral smear: 0% - 2% (adults)

15. BASOPHIL

16. BASOPHILS DISORDERS
• Basophilic granulocytes take part in the reactions of having the role of
degrading lipids.
• Basophilic granules contain histamine and heparin.
• Under the influence of histamine release factors, basophilic granulocytes
release histamine responsible for numerous allergic reactions.
• Basophilic granulocytes release all stored heparin during hyperlipemia
after food intake, which enables the degradation of triglycerides into
fatty acids and glycerol (heparin activates lipoproteinic lipase)
• Basopenia can occur as a response to thyrotoxicosis, acute
hypersensitivity reactions, and infections
• Basophilia can occur in people with hypothyroidism
• In the myeloproliferative disorders (e.g., polycythemia vera and
myelofibrosis), a marked increase in the number of basophils can occur

18. Monocyte
• Monocyte is round or oval shape
• Due to ameboid movements on the edge of the cell, 1, 2 or
more pseudopods can be seen
• The size is 12-20 μm
• N:C 2:1 – 1:1
• The cytoplasm is relatively abundant dull grey-blue
It may have a ground-glass appearance with a number of
evenly distributed fine azurophilic granules. Vacuoles
common
• The nucleus is usually round or kidney-shaped or horse-shoe
shaped but may be markedly indented or even lobulated,
with two or more lobes
• Nucleoli are not usually visible
• Bone Marrow: 2%
• Peripheral Smear: 2% - 9% (adults)

19. Monocyte

20. MONOCYTE DISORDERS
Monocytes help other white blood cells remove
dead or damaged tissues, destroy cancer cells,
and regulate immunity against foreign
substances.
After a few hours in the bloodstream, they
migrate to tissues (such as spleen, liver, lung, and
bone marrow tissue), where they mature into
macrophages as the main scavenger cells of the
immune system

21. MONOCYTE DISORDERS
• Monocytosis occurs in response to chronic
infections, in autoimmune disorders, in blood
disorders, cancers and in sarcoidosis (an
infection of the lung)
• Monocytopenia can occur in response to the
release of toxins into the blood by certain
types of bacteria as, well as in people
receiving chemotherapy or corticosteroids

23. LYMPHOCYTE
• Two types of lymphocytes can be distinguished:
- Small lymphocyte
- Large lymphocyte

24. SMALL LYMPHOCYTE
• Small lymphocytes are the major part of lymphocytes in the blood.
• The cell has a large roundish and regular nucleus.
• Size: about 9 - 12 μm
• N:C: 4:1
• Chromatin: Nucleus is eccentrically located, round or oval, often with
an indentation in one place; chromatin is heavily clumped, forming
compact lumpy accumulations without sharp demarcations
• Nucleoli not present
• Cytoplasm: Scanty cytoplasm is often barely visible and quite often
seen only at the nuclear periphery
No granules. Distinctly basophilic
• Bone Marrow: 5% - 15%
• Peripheral smear: 20% - 44% (adults)

25. SMALL LYMPHOCYTE

26. LARGE LYMPHOCYTE
• Large lymphocytes are the minor part of lymphocytes in the
bloodstream.
• The cell has a large roundish and regular nucleus
• Size: 9 to 18 μm
• N:C: 3:1
• Chromatin: Nucleus is slightly eccentric located, round or
oval, sometimes slightly indented
• Chromatin has a looser pattern and more transparent.
• Nucleoli is not present
• Cytoplasm: Larger amount of cytoplasm, sky-blue color
• There may be up to 10 azurophilic granules
• Distinguishing characteristic: Cytoplasm is more abundant
with tendency for azurophilic granules

27. LARGE LYMPHOCYTE

28. LYMPHOCYTES DISORDERS
• T- and B-lymphocytes have different roles in generating
acquired immunity:
- T-lymphocytes in cellular
- B-lymphocytes in humoral immunity
• Both types produce antibodies
• Together with natural killer-cells (NK-cells) and the
complement system they compose a powerful defensive
system in the organism
• Lymphocytopenia is an abnormally low lymphocyte count in
the body
• A reduction in the number of lymphocytes may not cause a
significant decrease in the total number of white blood cells

29. LYMPHOCYTOPENIA
• Viral infections – including HIV infection
• Lymphocytes can decrease briefly during
starvation, times of severe stress, and during
use of corticosteroids, chemotherapy for
cancer, and radiation therapy
• Cancer – leukemias, lymphomas
• Rheumatoid Arthritis
• Systemic Lupus Erthematosus (SLE)

30. LYMPHOCYTOSIS
Lymphocytosis is an abnormally high lymphocyte count in
the blood
The number of lymphocytes can increase in response to
infections, especially by viruses
Some bacterial infections, such as tuberculosis, may also
increase the number.
Certain types of cancer, such as lymphomas and acute or
chronic lymphocytic leukemia, may produce an increase in
the number of lymphocytes, in part by releasing immature
lymphocytes (lymphoblasts) or the lymphoma cells into the
bloodstream.
Graves' disease and Crohn's disease may also result in an
increase in the number of lymphocytes in the bloodstream

32. PLASMA CELL
• Size: 8 to 20 μm
• N:C: 2-1:1
• Nucleoli: None
• Nucleus: Round or oval, eccentric
• Chromatin: coarse
• Cytoplasm: Deeply basophilic, often with perinuclear zone (hof)
• Vacuoles: None to several
• Bone Marrow: 0% - 1%
• Peripheral blood: 0%
• Function of plasma cells is the synthesis and excretion of
immunoglobulins (antibodies)
• Plasma cells may be found in peripheral blood of patients with
measles, chickenpox, scarlet fever, multiple myeloma and plasmacytic
leukemia

33. PLASMA CELL

34. PLASMA CELL

35. REFERENCE RANGE
Cell 0- 4 years (%) Adult (%)
• NNeeuuttrroopphhiillss 2200--4444 5500--7700
• BBaannddss 00--55 22--66
• LLyymmpphhooccyytteess 4488--7788 2200--4444
• MMoonnooccyytteess 22--1111 22--99
• EEoossiinnoopphhiillss 11--44 00--44
• BBaassoopphhiillss 00--22 00--22

36. WHITE BLOOD CELLS

37. RELATIVE VS ABSOLUTE COUNT
• RReellaattiivvee aanndd aabbssoolluuttee counts are terms referring to the white cell
differential.
• The relative count refers to the percentage of a particular cell
counted from the 100 WBC differential
• The absolute count refers to the count derived from the total
white count multiplied by the percentage of any particular white
cell.
• The absolute count is clinically significant in many disease states
providing important information to the clinician.
• It provides a more accurate measure of the actual numbers of
cells present in the peripheral blood
• For example, Neutropenia in a patient, will place the patient at a
higher risk for infections. Patients undergoing chemotherapy will
need to get an absolute neutrophil count to monitor their
immune status

38. RELATIVE VS ABSOLUTE COUNT
Example:
• If the WBC is 5.0 x 103/μL
• And the differential reads:
- Segmented neutrophils: 40% (Range: 50% to 70%)
- Bands: 3% (Range: 2% to 6%)
- Lymphocytes: 55% (Range: 20% to 44%)
- Monocytes: 2% (Range: 2% to 9%)
• Then the absolute count of lymphocytes would be :
5000 X 0.55 = 2750
OR: (5 x 55) X 10 = 2750 OR: (5000 x 55) / 100 = 2750
• Absolute Neutrophil count = (% of Neutrophils + Bands) X
total WBC

39. RELATIVE VS ABSOLUTE COUNT
Normal values
• Neutrophils: 1800 – 6800 /μL
• Band Neutrophils: 0 – 700 /μL
• Lymphocytes: 1000 – 3400 /μL
• Monocytes: 200 – 800 /μL
• Eosionphils: 0 – 400 /μL
• Basophils: 0 – 100 /μL

40. BLOOD CELL COUNTING
THE END