seminar presentation

1. Morphology of Normal White
Blood Cells and Platelets
Presented by: Dr Prince Lokwani
Guided by: Dr.Poonam Mishra

2. WBC Normal values:
• WBC Count = 4,000 – 10,000/cu mm or
4 – 10 x 109
/L
• Differential Count:
• Neutrophil = 40 – 80 %
Segmenter = 35– 75 % ; Stab = 0 – 5 %
• Eosinophil = 1 – 6 %
• Basophil = <1-2 %
• Lymphocytes = 20 – 40 %
• Monocytes = 2 – 10 %

4. White Blood Cells
(Leukocytes)
Granulocytes
( segmented)
Agranulocytes
(nonsegmented)
Basophil
Eosinophil
Neutrophil
Monocyte
lymphocyte

5. Neutrophils
• PMN-Polymorphonuclear
Leucocytes.
• Appearance: pink granules
in cytoplasm, nucleus has 3-
4 lobes
• Function: Phagocytosis of
bacteria
• Azurophilic (1°) granules
are "lysosomes of PMNs",
occur in all leukocytes.

6. Band Cells (M3)
• Last immature stage in
Neutrophilic series
• Sometimes seen in
circulation
– Particularly during states
of chronic
infection,pregnancy
• Nucleus is elongated and of
uniform width

7. Metamyelocyte (M2)
• First stage that is clearly
divided into separate
lines
• Few hundred granules
present in the cytoplasm
• Specific granules
outnumber the azurophlic
granules 4:1
• Nucleus
– Heterochromatic
– Indentation deepens
to form horse-shoe
Late neutrophilic
metamylocyte
Neutrophilic metamylocytes
Eosinophil
metamylocyte

8. Myelocyte (M1)
• Spherical nucleus
– Becomes
increasingly
heterochromatic
• Prominent Golgi
apparatus
– Negative image
– Lots of azurophilic
granules
• Formation of specific
granules
– Emerge from Golgi
(cis face) complex
– Characteristic
staining reactions
for each line
• Last stage that can do
mitosis
Late Myelocyte/ Early Metamyelocyte
Neutrophil
myelocyte
Eosinophil
myelocyte
golgi

9. Promyelocyte
• First recognizable cell in
granulopoiesis(Cannot tell
what kind of cell it will
become)
• 17-26 um in diameter
• Largest cell in series
• Large oval nucleus
• Muliple nucleoli
• Golgi Ghost
• Azurophilic (primary)
granules in cytoplasm
– Produced only at this
stage
nucleoli
Azurophilic
granuels

10. myeloblast
• 15-20 µm
• large, euchromatic,
spherical nucleus
(1-4 nucleoli)
• basophilic cytoplasm
with no granules
• prominent nucleoli
• can be seen in
peripheral blood with
certain leukemias

11. Neutrophilia
(ANC>7,500/cmm)
• Infections(esp pyogenic), Inflammation,
Metabolic disorders
• Acute hemorrhage, corticosteroids
• Stress, post-surgery, burns, HDN
• Lithium drugs, neoplasms,smoking.

12. Neutropenia
(<2000/cmm)
• Decreased production
- Inherited/acquired stem cell disorder
- Benzene toxicity, cytotoxic drugs
• Increased destruction
- Immune mechanism, sequestration
• BM depression, IM, varicella, Typhoid
• SLE, hepatitis or any viral infections

13. Barr Body
• Sex chromatin
• Represents the second X chromosome
in females (2-3% of neutrophils in
females)
• Small,well-defined,round projection of

14. Vacuolated neutrophil
• Degeneration of cytoplasm begins to
acquire holes or as result of active
phagocytosis
• May reflect increased lysosomal
activity
• Found in: septicemia

15. ABNORMALITIES OF
LEUKOCYTES

16. Quantitative abnormalities
• Leucocytosis – substantial increase in the WBC
count.
- Physiologic increase (no trauma/injury)
- Pathologic increase (trauma/pathology)
• Leucopenia – substantial decrease in the WBC
count.
• N.V. = 4,000 – 10,000/cu mm

17. Eosinophil (Eos)
• Bilobed nucleus
• 1-6% of WBC
• Recruited to sites of
inflammation
• Function: Involved in
allergy, parasitic infections.
• Contains: Eosinophilic
granules
• Granules contain: Major
basic protein
Azurophilic granuels

18. Basophil
• Circulating form of mast
cells
• <1-2% WBC
• Contains: basophilic
granules
• Granules contain:
histamine and heparin
• IgE receptors
• Involved in allergy

19. Monocyte/ Macrophage
Monocyte
• 2-10% WBC
• Circulating form
(precursor) of tissue
macrophages
• Recruited to sites of
inflammation
Macrophages
• Phagocytosis, bacterial
killing, antigen
presentation
• Peritoneal cavity: peritoneal
macrophages
• Lung: alveolar macrophages
• Spleen: splenic
macrophages

20. lymphocyte
•Appearance: small (same size as
RBCs), little visible cytoplasm
•NO specific granules
• 20-40% of WBC
•T cells: CMI (for viral infections)
• B cells: humoral (antibody)
• Natural Killer Cells

21. Agranulocytes
1- Lymphocyte :
• Small & Large lymphocytes.
• Nucleus occupy most size of the cell,
leaving thin rim of cytoplasm.
• chromatin is of condensed type ( dark
stained)

22. 2- Monocyte :
• Large size cell
• Kidney shape or irregular nucleus
• Chromatin of the nucleus has thready appearance.
• Cytoplasm contain lysozymes ( cytoplasmic vacule)s).

23. Quantitative abnormalities
• Leucocytosis – substantial increase in the WBC
count.
- Physiologic increase (no trauma/injury)
- Pathologic increase (trauma/pathology)
• Leucopenia – substantial decrease in the WBC
count.
• N.V. = 4,000 – 10,000/cu mm

24. Neutrophilia
(ANC>7,500/cmm)
• Infections(esp pyogenic), Inflammation,
Metabolic disorders
• Acute hemorrhage, corticosteroids
• Stress, post-surgery, burns, HDN
• Lithium drugs, neoplasms,smoking.

25. Neutropenia
(<2000/cmm)
• Decreased production
- Inherited/acquired stem cell disorder
- Benzene toxicity, cytotoxic drugs
• Increased destruction
- Immune mechanism, sequestration
• BM depression, IM, varicella, Typhoid
• SLE, hepatitis or any viral infections

26. Eosinophilia
(AEC > 600/cmm)
• Allergic disorders (asthma)
• Parasitic infections (nematodes)
• Skin disease (eczema)
• Hodgkin’s disease
• Scarlet Fever
• Pernicious anemia

27. Eosinopenia
(< 50/cmm)
• Stress due to trauma or shock
• Mental distress
• Cushing’s syndrome
• ACTH administration

28. Basophil
BASOPHILIA
(>100/cmm)
Chronic myelocyic leukemia
Polycythemia vera
Hodgkin’s disease
BASOPENIA Hyperthyroidism
Pregnancy

29. Lymphocytosis
(ALC > 4000/cmm)
• Viral infections ( German measles )
• Infectious Mononucleosis (kissing dis.)
• Mumps (parotitis), pertussis
• Tuberculosis, syphilis, thyrotoxicosis

30. Lymphocytopenia(<1500/cmm)
• Congestive heart failure, SLE
• Renal failure
• Advanced Tuberculosis
• High levels of adrenal corticosteroids

31. Monocytosis
(AMC > 1000/cmm)
• SBE, Syphilis, Tuberculosis
• Protozoan infections
• Mycotic or fungal infections
• Malaria, Systemic lupus erythematosus
• Rheumatoid arthritis

32. Monocytopenia
• Lymphocytic leukemia
• Aplastic anemia

33. QUALITATIVE CHANGES-WBC
• Morphologic abnormalities involving either
the nucleus or cytoplasm
• Functional abnormalities
• Inherited or Acquired
• Examination of peripheral blood or a bone
marrow evaluation.

34. Abnormal granulocyte
morphology (inherited)
• Alder-Reilly anomaly - dense azurophilic
granules, mucopolysaccharoidoses
• May-Hegglin anomaly - Giant platelets,
Dohle-bodies like inclusions seen even in
monocytes
• Pelger Huet anomaly – failure of normal
segmentation of nucleus, bi-lobed nucleus
or stab forms only,
“pince-nez nucleus”

35. Alder-Reilly anomaly
• Heavy,coarse blue-
black granules of
BEN & sometimes
lymphocytes &
monocytes
• Inherited condition
• Associated with
Hurler’s syndrome &
Hunter’s syndrome

36. May-Hegglin Anomaly
• Benign inherited anomaly affecting all
leucocytes except lymphocytes.
• Larger than usual Dohle-like bodies

37. Pelger-Huet Anomaly
• Indicates failure of
neutrophil to segment
properly
• Bi-lobed nucleus;
chromatin is coarsely
clumped
• May be inherited or
acquired (as in
leukemias)
• Heterozygous for this
char.shows numerous bi-
lobed (dumbell shape);
homozygous-round
neutrophil

38. Continuation:
• Chediak Steinbrinck Higashi syndrome – large
lysosomes containing hydrolases and other enzymes.
There is anemia,thrombocytopenia, leucopenia and
increased susceptibility to infection. There is partial
albinism & photophobia.
• Also seen in Aleutian mink, mice, cat, cattle & killer
whale as caused by abnormal WBCs.

39. Chediak-Higashi Syndrome
(Autosomal recessive disorder)• Rare,fatal disprder found in
children
• Inherited as an autosomal
recessive char.
• Contain very large,reddish-
purple or greenish-gray
staining granules in the
cytoplasm of granulocytes
• In monocytes & lymphocytes,
stain bluish-purple
• These granules represent
abnormal lysosomes
• Found in: anemia
neutropenia
thrombocytopenia

40. QUALITATIVE CHANGES-WBC
• Morphologic abnormalities involving either
the nucleus or cytoplasm
• Functional abnormalities
• Inherited or Acquired
• Examination of peripheral blood or a bone
marrow evaluation

41. Abnormal granulocyte
morphology (acquired)
• Cytoplasmic abnormalities
• Nuclear abnormalities

42. • Dohle bodies,
Diffuse basophilia
• Cytoplasmic foaminess
• Toxic granulation
Cytoplasmic abnormalities

43. Dohle Bodies
• The earliest and first indication of toxic change.
• Single or multiple light blue or gray areas in cytoplasm of
neutrophils
• RER & represent failure of cytoplasm to mature
• Found in: infections, poisoning, burns, following
chemotherapy

44. Diffuse cytoplasmic basophilia
• diffuse irregular blue appearance to the
cytoplasm.
• Due to the presence of polyribosomes and
rough endoplasmic reticulum.
• Can be seen during bacteremia and
generalized infection .

45. Foamy cell
• These are indistinct vacuoles in the
cytoplasm, giving it a frothy appearance
• Due to degranulation of lysosomes, which
result in autodigesion.
• Appear in sever inflammation.

46. Toxic granulation
• Characterized by prominent purblish cytoplasmic
granules.
• 1ry
granules retain their staining affenity.
• most commonly seen in large animals (horses, ruminants,
camelids ).
• it’s presence suggest severe inflammatory process.

47. Nuclear abnormalities
• Band cell
• Hypersegmentation
• Giant hypersegmentation

48. Band neutrophil
Hypersegmented
neutrophil
(>5 segments)
Normal neutrophil
(3-5 segments)
Shift to left
(Inflammation)
Shift to right
(Aging)

49. Giant hypersegmentation
• Occurs in vitamin B12 and folic acid
deficiency (Megaloblastic anemia)

50. Other abnormalities:
• Smudge or basket cells – squash-degenerated
nucleus of WBCs
• Jordan’s anomaly – fat-containing vacuoles in
WBC cytoplasm, Ichthyosis
• Twinning deformity
• Auer rod – rod-like structure seen in the
cytoplasm of myeloblasts, diagnostic for Acute
myeloblastic leukemia (AML)

51. Reactive lymphocytes

52. Downey cell
• Hallmark cell seen in cases of Infectious
mononucleosis (kissing disease)
• Atypical lymphocyte (stress lymphocyte)
• “ballerina skirt cell”

53. Infectious Mononucleosis

54. Plasma cells
• Ovoid or fibrillary shaped
• Eccentric location of nucleus
• Perinuclear halo
• “cart-wheel pattern or spoke of the wheel
pattern of nucleus”
• basophilic cytoplasm

55. Inherited abnormalities involving
Monocyte-macrophage group
• MUCOPOLYSACCHAROIDOSES
- Hunter syndrome, Hurler’s disease
• LIPIDOSES – lipid accumulation
- Gaucher’s disease – accumulation of
glucocerebroside due to lack of beta-
glucosidase enzyme
- Neimann Pick disease – sphingomyelin and
cholesterol accumulation due to lack of the
enzyme sphingomyelinase

56. Monocyte-macrophage abnormality

57. Leukemias
• In the case of WBC, these malignant cells may or
may not circulate in the peripheral blood. Hence,
WBC count may be increased or otherwise.
• Should these abnormal cells be present both in
the bone marrow and the peripheral blood, the
term leukemia is used.
• Aleukemic leukemia – if only confined to the
marrow and do not circulate.

58. Classification of the leukemias
• According to the stem cell line involved
- Myeloid – involves the granulocytes,
monocytes, RBCs and megakaryocytes. Also
known as myeloproliferative disorders or
nonlymphocytic leukemias.
- Lymphoid – involving the B or T cells and may
be a leukemia or lymphoma

59. Classification of the leukemias
• According to the stem cell line involved
- Myeloid – involves the granulocytes,
monocytes, RBCs and megakaryocytes. Also
known as myeloproliferative disorders or
nonlymphocytic leukemias.
- Lymphoid – involving the B or T cells and may
be a leukemia or lymphoma

60. Classification of leukemias
• According to duration (life span)
- Acute – days to weeks (3 months)
- greater than 30 % blasts forms
- Chronic – more than a year (1-2 years)
- less than 10 % blast forms

61. Examples :
Acute myeloid leukemia myeloblast
Chronic myelogenous
leukemia
Myelocyte, metamyelocyte &
neutro
Acute lymphoblastic
leukemia
lymphoblasts
Chronic lymphocytic
leukemia
Small mature lymph
Erythroleukemia
Di Guglielmo syndrome
> 50% of the nucleated cells
are erythroblasts

62. Comments on the leukemias:
• AML – most common form of acute leukemias
in first few months of life, in middle aged
group and later years
• CML – more common in young & elders
• ALL – seen among children 2 – 10 y.o.
• CLL – common among > 60 years old

63. Overview of Disorders of
Platelets

64. STRUTURE
Mucopolysacch.
coat
αGranules
Dense core
granules
Mucopolysacch. Coat: Glycoprotein content which are
important for interaction of platelets with each other
or aggregating agents.
− α Granules:
- Dense core:

65. Platelets (*)
• (*) are released from the megakaryocytes, likely
under the influence of flow in the capillary sinuses.
• Main regulator of (*)production is the hormone
thrombopoietin (TPO), which is synthesized in the
liver.
• Normal BLOOD platelet count = 150,000–450,000/L.
• (*) synthesis increases with inflammation and
specifically by interleukin 6.

66. Platelets
• Are very active, aneucleate and they have limited
capacity to synthesize new proteins
• Circulate with an average life span of 7–10 days.
• Approximately 1/3 of the platelets reside in the
spleen, and this number increases in proportion to
splenic size, although the platelet count rarely
decreases to <40,000/L as the spleen enlarges.

68. Thank You For Your Attention