Download Granulocytes

Blood
160 SFST
Blood
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Blood smear
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Differential count
Bone marrow smear
Blood
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Volume – approximately 5,5 L
Hematocrit – volume of erythrocytes
(percentage): 35 – 45% in women, 40 – 50% in
men
1% leucocytes and platelets (buffy coat)
Rest – plasma (water, inorganic salts, organic
compounds, proteins (albumin, alfa, beta, gama
globulins, lipoproteins, fibrinogen)
Serum – after removal of fibrinogen and
coagulation factors
Blood smear
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Droplet of peripheral blood on
the slide
Dry, then stain according to
Pappenheim:
3 minutes by May-Grünwald
solution (fixation by methanol),
solve and stain for 1 minute.
Then stain by GiemsaRomanovski solution for 15 - 20
minutes. Wash and dry.
Observe under immersion
objective.
Blood smear
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Staining according to Pappenheim:
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May-Grünwald–
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Giemsa-Romanovski
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Eosin-methylene blue - 1 g
Methanol - 100 ml
Glycerol - 50 ml
Azur- eosin II - 3 g
Azur II - 0,8 g
Glycerol - 250 g
Methanol - 250g
Result is similar to the hematoxylin eosin staining
Erythrocytes
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Biconcave discs without nuclei
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Size: 7,5 x 2,6 μm (in the middle only 0,8 μm)
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Number: 3,9 – 5,5 milions in 1μL
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Plasmalemma, cytoskeleton, hemoglobin
(33%), enzymes: glycolysis, hexosemonophosphate shunt
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They survive 120 days in circulation
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Function: transport of gases (O2, CO2)
Neutrophilic granulocyte
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60-70%
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Size: 12-15 μm (in smear)
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Nucleus is divided in two or three lobes that are connected
by chromatin briges; inactive X chromosome – Barr body
Specific granules – small, near the limit of resolution in SM
- lysosomes
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Azurophilic granules – 0,5 μm
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Glycogen – source of energy – anaerobic metabolism
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Function: Nonspecific immunity: phagocytosis, oxidative
burst – (H2O2) and oxygen radicals
Neutrophilic granulocyte
Eosinophilic granulocyte
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2-4%
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Size: 12-15 μm
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Bilobed nucleus
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Specific granules: crystalline core (internum) – major
basic protein
matrix (externum) - lighter
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Function: activity again parasitic infections
Modulation of inflamation (inactivation of leukotrienes and
histamine)
Eosinophilia – typical for parasitic infection, alergy
Eosinophilic granulocyte
Basophilic granulocyte
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Less that 1%
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Size 12-15 μm
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Nucleus is divided in irregular lobes, but it is not
distinct because it is covered by granules
Specific granules – metachromatic, content of
heparine and histamine – liberation of granules
– degranulation - after binding of certain
antigens.
Function – modulation of immune response
Basophilic granulocyte
Lymphocyte
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Division according to the size: small (6-8μm),
medium-sized and large (up to 18μm).
Division according to the specific superficial
antigens – B, T, NK
Small lymphocytes prevail in blood – memory
cells
Large round nucleus, chromatin is condensed,
nucleolus
Thin rim of cytoplasm, ribosomes, azurophilic
granules
Function
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T and B lymphocytes – specific immunity (T
lymphocytes prevail in peripheral blood: 65% 75% )
B lymphocytes – humoral – differentiation in
plasma cells – production of antibodies
T lymphocytes – cytotoxic CD8, helper CD4
NK (Natural killers) lymphocytes – 10-15% in
peripheral blood – nonspecific - innate immune
response (induce apoptosis)
Lymphocyte
Monocyte
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Size: 12 -20μm
Oval, horseshoe or kidney shaped nucleus,
excentrically placed
Basophilic cytoplasm, azurophilic granules
(lysosomes), RER, polyribosomes,
mitochondria, Golgi complex
Monocytes differentiate into macrophages
(antigen presenting cells)
Function: phagocytosis, antigen-presenting
cells
Monocyte
Platelet
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Nonnucleated, disclike cell fragments of megakaryocytes
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Size - 2-4 μm
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200 000– 400 000 in uL
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Central zone containing granules – granulomera and
peripheral lighter zone - hyalomera
Hyalomera: open canalicular system – invagination of
superficial plasmalemma, marginal bundle of microtubules,
actin and myosin – contractile
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Granulomera: Mitochondria, glycogen, granules
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Function: coagulation
Platelets
Leukocytes
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6 000 -10 000 in 1μL
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Differencial count:
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Granulocytes
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Neutrophiles 60 -70%
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Eosinophiles 2-4%
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Basophiles
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Agranulocytes
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Lymphocytes 20-30%
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Monocytes
0-1%
3-8%
Neutrophilic granulocytes
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2,7 – mean of nuclear segments in neutrophilic
granulocytes
Shift to the left – release of younger neutrophilic
granulocytes – sign of the inflamation
Shift to the right – more segments – failure of
neutrophilic granulocyte development
(hemopoiesis) – absence of vitamines
Red bone marrow
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Stroma – reticular
connective tissue
(reticular cells and fibres
– collagen type 1 and 3,
fibronectin, laminin,
proteoglycans)
Sinusoids – capillaries
with discontinuous
endothelium
Red bone marrow
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Stroma: reticular connective tissue – reticular
cells and extracellular matrix
Capillaries with discontinuous endothelium
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Adipocytes
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Stem cells
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Erythroblastic islands
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Myelopoetic cells
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Megakaryocytes
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Mast cells
Hemopoiesis
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Erythrocyte development:
Proteosyntesis – syntesis of
hemoglobin
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Loss of nucleus and ribosomes
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Proerythroblast
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Formation of azurophilic
and specific granules
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Segmentation of nucleus
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Myeloblast
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Promyelocyte
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Myelocyte
Basophilic erythroblast
Polychromatophilic erythroblast
Orthochromatophilic
erythroblast
Granulocyte
development:
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Reticulocyte
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Metamyelocyte
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Erythrocyte
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Mature leukocyte
Hemopoiesis
Erythroblastic island