Histology of blood

Ma. Minda Luz M. Manuguid M.D.

 blood
 def.: specialized, liquid connective
tissue ubiquitous in the human body,
circulating in a closed system of
vessels
Blood cells: Erythrocytes,
Leukocytes, Thrombocytes
Ground substance: Plasma
Extracellular fibers: potential
(Fibrinogen) become actual fibers
(Fibrin) when blood clots
 Blood cells
Erythrocytes – red blood cells
Leukocytes – white blood cells
Granulocytes
• Neutrophils
• Eosinophils
• Basophils
Agranulocytes
• Lymphocytes
• Monocytes
Thrombocytes – platelets
 Erythrocytes
 most prevalent of
blood cells: 25 trillion
in p.b.; comprise 45%
of total blood volume
 anucleate biconcave
discs 7-8µm in
diameter, 2µm thick
at the edges, 1µm at
the center; bound by
plasma membrane
 Hemoglobin is 1/3 of
rbc mass
 cytoskeletal proteins:
Erythrocytes (RBC, Red corpuscles)
Red blood cells
 no nucleus
 cytoplasm is entirely
acidophilic (red)
 biconcave shape,
 6.5 to 8.4μ in diameter
 indention or notches
and minor surface
irregularities are
considered normal
variations. 
 erythropoiesis
CFU-S → BFU-E → CFU-E EPO
→→→
Erythroblast
colony burst colony
↓
forming forming forming
basophilic
unit-Spleen unit-Erythroid unit-Erythroid
Erythroblast
↓
polychromatophilic
EPO: Erythropoietin
Erythroblast
↓
orthochromatic
Erythroblast
 erythroblast – proNormoblast
 basophilic erythroblast – early Normoblast
 polychromatophilic erythroblast –
intermediate Normoblast
 orthochromatic erythroblast – late
Normoblast
 Rbc maturation
 Immature cells have delicate, fine nuclear
chromatin which gradually becomes coarsely
clumped or condensed. The size of the nucleus
decreases; nucleoli are reduced in number or lost
completely as in red cells. The nuclear shape
which is initially round or oval may become
uniquely configured as in myeloid cells. Mitotic
competence is lost as cells differentiate.
 Rbc maturation
 although most erythroid and myeloid cells
decrease in size as they mature, the
cytoplasm becomes relatively increased in
volume, compared to the nucleus. The
cytoplasm becomes less basophilic and
increasingly eosinophilic.
 Pronormoblast / erythroblast
 The pronormoblast
(E1) is the earilest
recognizable red
cell precursor.
They are 15- 20µ in
diameter with fine
granular chromatin
and 0-2 nucleoli.
The cytoplasm is
deeply basophilic
without granules.
 Basophilic erythroblasts
Basophilic normoblasts:
 E2:The chromatin is
slightly coarser and
the cell slightly
smaller than the
pronormoblast.
Nucleoli may be
indistinct. The
cytoplasm remains
deeply basophilic.
 E3 : The chromatin
becomes increasingly
coarse compared to
the E2 stage. The
cytoplasm remains
 Polychromatophilic erythroblasts

 E4:
polychromatophilic
normoblasts:
Hemoglobin
production
becomes evident
from the muddy
mix of the blue-
staining RNA and
the red-staining
hemoglobin
resulting in the
 Orthochromatic erythroblast
 The E5
orthochromatic
normoblast is
smaller than the E4
stage.
 hemoglobin is
abundant while
ribosomes
decrease in
number.
 The nuclear
chromatin is
 reticulocytes
 Polychromatophilic red
cells are larger than
mature erythrocytes
and still contain
ribosomes and
mitochondria, giving
the cell a blue tint.
When polychromatic
RBCs are stained with
a supravital dye (new
methylene blue or
cresyl violet)
ribosomes clump to
form a blue stained
 Reticulocytes & erythrocytes

Reticulocyte –
immature rbc
with aggregated
reticular clusters
of ribosomes
 used as marker
for bone marrow
hematopoiesis
e.g. in treating
anemia
 erythrocytes
 Normal mature
erythrocytes are
biconcave discs
(approximately 8
um in diameter;
2.5 um thick at the
periphery; 1.0 um
thick at the
center). This
results in an area
of central pallor
(about 1/3 the dia.)
Erythroid hyperplasia & normal
marrow
Granulocytes
 Leukocytes: Myeloid series

 the mature forms of the myeloid series

(neutrophils, eosinophils, basophils), all
have lobed (segmented) nuclei. The
degree of lobation increases as the cells
mature.
 myeloblast
 earliest recognizable
myeloid cell is the
myeloblast (10-20m
dia) with a large round
to oval nucleus. There is
fine diffuse immature
chromatin (without
clumping) and a
prominant nucleolus.
 The cytoplasm is
basophilic without
granules. Although one
may see a small golgi
area adjacent to the
nucleus, granules are
 promyelocyte
 promyelocyte (10-
20m) is slightly larger
than a blast. Its
nucleus, although
similar to a myeloblast
shows slight
chromatin
condensation and less
prominent nucleoli.
 The cytoplasm
contains striking
azurophilic granules or
primary granules.
These granules
contain
myeloperoxidase, acid
phosphatase, and
 myelocyte
 when secondary granules
can be recognized, the cell
becomes a myelocyte.
 slightly smaller than
promyelocytes, have
eccentric round-oval nuclei,
often flattened along one
side. The chromatin is fine,
but shows evidence of
condensation. Nucleoli may
be seen in early stages but
not in the late myelocyte.
 Primary azurophilic granules
are still present, but
secondary granules
predominate. Secondary
granules (neut, eos, or baso)
first appear adjacent to the
 metamyelocyte
 Metamyelocytes (10-
18m) are slightly
smaller than
myelocytes.
 They have kidney
shaped indented
nuclei and relatively
dense chromatin,
especially along the
nuclear membrane.
 The cytoplasm is
faintly pink with
almost no blue
background.
Numerous secondary
granules (neutro, eos,
or baso) clearly
 bands
 Bands, slightly smaller
than juveniles, are
marked by a U-shaped
or deeply indented
nucleus. Opposite
sides or lobes are of
roughly equal size or
diameter. There is no
nuclear constriction >
than 1/2 the lobe
diameter. The
chromatin is heavily
clumped and
secondary or specific
granules either
neutrophilic or
basophilic
predominate.
 neutrophils
 Segmented (segs) or
polymorphonuclear (PMN)
leukocytes (average 14 m
dia) are distinguished by
definite lobation with thin
thread-like filaments of
chromatin joining the 2-5
lobes. The chromatin of the
segmented neutrophil is
coarsely clumped and the
cytoplasm is pink due to
large numbers of secondary
granules.
 In practice when examining
peripheral blood, neutrophils
are the only leukocytes to be
divided into myelocyte,
juvenile, band, and PMN
stages. Eosinophils and
 neutrophils
 the mature neutrophil
(12-15u dia) is
characterized by
segmentation into 2-5
lobes. The chromatin
is dense and clumped
with distinct lighter
areas of
parachromatin.
 The cytoplasm is
lightly eosinophilic
with variable numbers
of light staining
"neutral" granules and
a few and a few
 eosinophils
 The large orange
granules of the
eosinophil make it the
most readily
recognizable cell in the
blood. The eosinophil
(12-15u diameter) has
chromatin similar to that
of a neutrophil, but
usually fewer (2-3) lobes.
 Normally 0-6%
eosinophils are found in
the peripheral blood.
 The eosinophil specific
granules contain
rhomboid crystals by EM.
This core contains Major
 basophil
 The purple-black,
often large coarse
irregularly sized
granules of the
basophil are
chacteristic and may
obscure the nucleus of
this relatively
uncommon cell.
 the mature nucleus is
segmented into 2-3
lobes.
 Basophils (12-15u dia)
contain heparin and
large amounts of
Agranulocytes
 monocytes
 the largest (12-20u dia)
cells normally found in the
peripheral blood.
 have abundant blue-gray,
sometimes very pale pink
cytoplasm with small,
generally indistinct,
granules. One may see
fine reddish granules of
variable prominence.
Occasionally, large
azurophilic granules will be
seen. Cytoplasmic
vacuoles are often seen.
 The nucleus is irregular,
 lymphocytes
 vary greatly in size (7-
20m dia) and in
nuclear and
cytoplasmic character.
 The small lymphocyte
(7-12m dia) has a
round-oval nucleus
with dense clumped
"smudgy" chromatin;
no visible nucleolus.
 Scanty light blue
cytoplasm, while
usually agranular,
may contain a few
 lymphocytes

 Lymphocytes with peripherially clumped

chromatin and often deep blue cytplasm
similar to plasma cells are termed
plasmacytoid lymphocytes.
 Normal peripheral blood lymphocyte
percentages vary from 15-60%.
 Plasma cell
 Terminally
differentiated
lymphocytes or
plasma cells (left
panel) are rarely
found in the
peripheral blood.
 platelets
 Immediately above
the leukocytes in the
buffy coat lies a thin
layer of platelets. On
Wright's stained
peripheral smear
platelets are small (2-
5m) round-oval
anuclear bits of pale
blue cytoplasm
containing azurophilic
granules.
 Smaller platelets tend
to be older; while
Thrombocytes
 megakaryocytes

 Megakaryocytes undergo endomitosis - a process

whereby DNA is duplicated without cell division.
Thus MKs become polyploid during amplification.
 Most platelet forming MKs have a ploidy of 16N,
(range 4N - 64N).
 MKs are the largest normally occuring cells in the
 megakaryocytes
 Closer inspection of
MKs shows a
network of platelet
demarcation zones
forming fissures
throughout the MK
cytoplasm. The
membranes of this
demarcation
system eventually
form cytoplasmic
platelet
megakaryocyte
 platelets

 Platelet granules are of two types: alpha granules

and dense bodies.
 Alpha granules contain Platelet Derived Growth
Factor, platelet factor 4, Factors V & XIII and
fibrinogen.
 Dense bodies contain serotonin,nucleotides (ADP)
and calcium. Lysosomes containing hydrolytic
 Thrombocytes / platelets
 The peripheral blood
platelet count ranges from
150 - 450 x 10 /L.
 About 2/3 of platelets
circulate, while 1/3 are in
the splenic pool or other
extravascular locations.
 During its lifespan the
average megakaryocyte
(MK) gives rise to
approximately 4,000
platelets which live an
average of 9-12 days.
 In the steady state, where
platelet production =
platelet destruction, daily
That’s all !