HISTOLOGY

BLOOD & HEMATOPOIESIS I

Blood: Specialized CT
All

connective tissue consists of cells embedded

in a matrix that consists of ground substance and
fibers.
Blood may thus be described as connective tissue
whose matrix consists of free-flowing ground
substance (plasma) with no fibers.

Blood: Specialized CT
All

of the several blood cell types originate in

the connective tissue of bone marrow.
Certain white blood cells, notably lymphocytes
and monocytes, move freely back and forth
between blood and other connective tissues.
The chemical composition of plasma is very
similar to that of interstitial fluid in ordinary
connective tissue.

Blood
Blood
Formed elements

Cells & Platelets

RBC = 44%
WBC = 1%
Plasma = 55%

Plasma is an aqueous solution containing

proteins (7%), lipoproteins (10%),
amino-acids, vitamins, hormones, and inorganic salts.
The main blood proteins are:
(a) albumins (maintains osmotic pressure of blood)
(b) gamma globulins (immunoglobulins or antibodies)
(c) fibrinogen (clotting agent)

Fibrinogen+

Serum

Suitable Slides
blood smear - Leishman, Wright's, Giemsa or May-Grnwald-Giemsa stains

Cellular components

Red blood cells

erythrocytes (44%)
White blood cells
(leukocytes) and platelets
(1%)

RED BLOOD CELLS erythrocytes

1. Biconcave discs about 7.2m diameter [2.6 Vs0.8]
(similar to the dimensions of most capillaries), nonnucleated,
2. Acidophilic and are stained a reddish color with eosin.
3. Main Protein is Hemoglobin + O = Oxyhemoglobin
4. 120 day life span
5. Blood count: males have more per unit volume
(5.5 x 106/ul of blood) than females (4.1-6) [3.9-5.5]

0.8

2.6

RED BLOOD CELLS erythrocytes

6. Carbonic anhydrase CO2 transportation
7. Enzymes of Glycolytic and HMP shunt pathways
8. ABO blood group system
9. Rh antigens Rh+ vs Rh

Type O blood is the most common blood type, followed by type A, type B, and, the least common blood type, AB.
Blood type

Antigen present

Blood type

Antigen

Antibody

anti-B

anti-A

AB

A& B

AB

A& B

neither

neither A nor B

neither

both anti-A & anti-B

Drop of blood in which

anti-A antibody was
added

Drop of blood in which

anti-B antibody was
added

Blood
type

Clumping

No clumping

No clumping

Clumping

Clumping

Clumping

AB

No clumping

No clumping

Type O:
universal donor
no antigens = no
clumping
Type AB:
universal recipient
no antibodies = no
clumping

Rh system: inherited independent of ABO system

Rh positive = antigen present on RBCs (& no
antibodies)
Rh negative = no antigen & antibodies will be produced
IF exposure occurs
Erythroblastosis fetalis (also called Rh disease):
hemolysis of RBCs of fetus which can cause anemia or
worse; may occur when an Rh negative mother & Rh
positive father have an Rh positive fetus
RhoGAM: treatment for Rh disease;
contains antibodies specific for

O+ 37%, O- 6%, A+ 34%, A- 6%,

B+ 10%, B- 2%, AB+ 4%, AB- 1%

Clinical Applications
1. Anemia: Abnormal low values of hemoglobin
2. Polycythemia or Erythrocytosis
3. Erythrocyte membrane deformation
cytoskeletal organization -Spectrin
4. Anisocytosis Abnormal variation in cell size
(macro and microcytosis)
5. Poikilocytosis altered cell shape (sickling, crenation)

Hemoglobin

composed of globin (made up

of 4 highly folded polypeptide
chains) + 4 heme groups (with
iron)
each molecule can carry 4
molecules of oxygen
called oxyhemoglobin when
carrying oxygen & called
reduced hemoglobin when not
carrying oxygen
can also combine with carbon
dioxide & helps transport
carbon dioxide from the tissues
to the lungs

WHITE BLOOD CELLS - leukocytes

Granulocytes

Agranulocytes

neutrophils, eosinophils and

basophils

lymphocytes and monocytes

~ 60% neutrophils (50% - 70%)

~ 3% eosinophils (>0% - 5%)
~ 0.5% basophils (>0% - 2%)
~ 5% monocytes (1% - 9%)
~ 30% lymphocytes (20% - 40%)

Granulocytes are characterized by:

(i) irregular segmented nuclei
(ii) specific granules (specific size, staining
affinities, ultrastructure)
(iii) terminal (fully differentiated) cells.
Agranulocytes are characterized by:
(i) regular nuclei (round or kidney-shaped)
(ii) non-specific granules (also present in other
leukocytes)

Processes exhibited by Leukocytes

Chemotaxis

Amoeboid movement

Chemical attraction which stimulates movement of WBC,

especially Neutrophils and monocytes, to sites of infection
WBC motility in which pseudopodia are extended and the rest of
the cell pulled forward in a crawling action along and through
the vascular wall.

Diapedesis

Name for the process in which WBCs flatten and move through
the vascular wall into the interstitial tissue

 Neutrophils

(also called neutrophilic

granulocytes, or polymorphonuclear
neutrophilic leukocytes, PMNs, or polys) are
the most numerous of the leukocytes, about
60% of the white blood cell count.
They are about 12 m in diameter in blood
smear preparations (about twice the size of red
blood cells

PMN Leukocytes

Neutrophil granulocytes (or neutrophils)

Have a very characteristic nucleus.

Divided into 3-5 lobes which are connected
by thin strands of chromatin.
The number of lobes increases with cell age.
Up to 7 lobes can be found in very old
neutrophils (hypersegmented cells).
Neutrophils also contain two types of granules.
Primary granules (or A granules) contain lysosomal enzymes and
are likely to be primary lysosomes, although they are larger (0.4
m) than the "ordinary" primary lysosome.
Secondary granules (or B granules), the specific granules of the
neutrophils, contain enzymes with strong bactericidal actions.

The specific granules of neutrophils stain only weakly if they are at all
visible - they are "neutral", hence the term neutrophil.

Function

Neutrophils play a central role in inflammatory processes.

Large numbers invade sites of infection in response to factors (e.g.
cytokines) released by cells which reside at an infection site.
Neutrophils are the first wave of cells invading infection sites.
Receptors in their plasma membrane allow them to recognize foreign
bodies, e.g. bacteria, and tissue debris, which they begin to
phagocytose and destroy.
The phagocytotic activity of neurophils is further stimulated if
invading microorganisms are "tagged" with antibodies (or opsonised).
Neutrophils cannot replenish their store of granules. The cells die
once their supply of granules has been exhausted. Dead neutrophils
and tissue debris are the major components of pus.
Their lifespan is only about one week.

EOSINOPHILS: 10 15 um
1.
2.

Typically with 2 lobes

1-4% of leukocytes (150 400 / ul of blood)

3. Short life few days

4.

Granules specific and nonspecific (azurophilic/acidophilic)

a. specific: large granules (crystalloid-major basic protein, enzymes)
b. nonspecific: lysosomes; hydrolytic enzymes

EOSINOPHILS: 10 15 um
Function: destroy antibodyantigen complexes and parasitic worms
. Release Major Basic Protein involved in killing parasites
. Engulf and destroy antibodyantigen complexes

BASOPHILS: 8 12 m
1. Less than 0.5-1% of leukocytes (50 100 / ul of blood)
2. Basophilic granulocytes have a 2 or 3 lobed nucleus. The lobes
are usually not as well defined as in neutrophilic granulocytes
and the nucleus may appear S-shaped.

The specific
granules of basophils are stained deeply bluish or reddish-violet.
4. granules specific and nonspecific (azurophilic)
a. specific: large granules (heparin, histamine, Leukotriene)
b. nonspecific: lysosomes; hydrolytic enzymes

Functions

Heparin and histamine are vasoactive substances. They dilate

the blood vessels, make vessel walls more permeable and
prevent blood coagulation.
As a consequence, they facilitate the access of other
lymphocytes and of plasma-borne substances of importance for
the immune response (e.g. antibodies) to e.g. a site of
infection.
The release of the contents of the granules of basophils is
receptor-mediated.
Antibodies produced by plasma cells (activated Blymphocytes) bind to Fc-receptors on the plasma membrane of
basophils.
If these antibodies come into contact with their antigens, they
induce the release of the contents of the basophil granules.

Remember mast cells?

1. Binding of antigens (to IgE) releases specific granules
2. Histamine vasodilation, smooth muscle contraction, etc.
3. Heparin anticoagulant

Size
Nuclear

config
Granule distrib
Granule content

MONOCYTES: 12 25 um
1. Kidney or C shaped nucleus; no specific granules;
"pale blue" cytoplasm
2. Life span few days in circulation / months in tissues
3. 3% of leukocytes (200 800 / ul of blood)
4. Granules: nonspecific (azurophilic) lysosomes

MONOCYTES: 12 25 um
Function is to become tissue macrophages after migration:
1. avid phagocytes (dead cells, antigens, bacteria)
2. produce cytokines initiate inflammation, etc.
3. antigenpresenting cells
4. foreign body giant cells
(multinucleated)

LYMPHOCYTES: 5 18 um
a. 25 30% of leukocytes (1500 2500 /ul of blood)
b. Life span months to years
c. Round, dense nucleus; no specific granules;
small, "blue" cytoplasm [may be large or small]
d. Granules: nonspecific (azurophilic) lysosomes

LYMPHOCYTES: 8 18 um
Function: immune response (B cells, T cells, null cells)
1. B-cells become plasma cells produce antibodies
2. T-cells kill viral cells, produce cytokines
3. Null cells stem cells, natural killer cells

PLATELETS: 2 4 um
No nucleus; never had one
2. Cell fragments from megakaryocytes
3. Hyalomere: outer region, hardly stains, peripheral tubular
system, increases surface area
4. Granulomere: dense core with granules and few organelles,
stains bluish
5. Granules: alpha, delta, lambda (lysosomes)
1.

a. alpha: fibrinogen, platelet derived growth factor,[PDGF], etc.

b. delta: calcium, serotonin, ATP, etc.
c. lambda: lysosomal (hydrolytic) enzymes

Count: 250,000 500,000

Platelets assist in hemostasis, the arrest of bleeding.

Serotonin is a potent vasoconstrictor. The release of serotonin from

thrombocytes, which adhere to the walls of a damaged vessels, is
sufficient to close even small arteries.

Platelets, which come into contact with collagenous fibers in the walls
of the vessel (which are not usually exposed to the blood stream),
swell, become "sticky" and activate other platelets to undergo the same
transformation. This cascade of events results in the formation of a
platelet plug (or platelet thrombus).

Finally, activating substances are released from the damaged vessel

walls and from the platelets. These substances mediate the conversion
of the plasma protein prothrombin into thrombin. Thrombin catalyzes
the conversion of fibrinogen into fibrin, which polymerizes into fibrils
and forms a fibrous net in the arising blood clot.

Platelets captured in the fibrin net contract leading to clot retraction,

which further assists in hemostasis.

PLATELET FUNCTION

Some Anomalies
Inappropriate clotting:
Thrombus - clot formed in an intact vessel, possibly due to:
roughened vessel walls (atherosclerosis)
slow-moving blood (e.g., in varicose veins) = small quantities of fibrin
form & accumulate
Embolus - 'moving' clot = piece breaks-off from a thrombus.
Excessive bleeding:
Hemophilia
genetic 'defect'
inability to produce certain factor(s)
Thrombocytopenia
abnormally low platelet count
most persons have idiopathic thrombocytopenia (= unknown cause) while
in others it's an autoimmune disease

Identify the cells, their size, their granules and their function:

Identify the cells, their size, their granules and their function:

Identify the cells, their size, their granules and their function:
A

NORMAL BLOOD CELL COUNT