Chapter 3Blood

1 Overview of Blood
Blood is a fluid connective tissue that circulates through the
cardiovascular system.
Blood consists of blood cells and extracellular protein-rich fluid called
plasma

Blood consists of a variety of cells suspended in a fluid medium called

plasma.
Blood is one kind of connective tissues.
It functions as a vehicle for the transport of gases, nutrients, metabolic
waste products, cells and hormones throughout the body.
(vehicle)

()

2Blood cell types

The cells of blood are of three major classes

Red blood cells

erythrocytes

X()
White blood cells leukocytes

Platelets
thrombocyte
X

s
They are formed in the hemopoitic tissues, bone marrow and lymphatic
tissue
(hemopoitic tissues)

3Histological methods used to study blood and bone

marrow

Blood Spread (smear)

1. 1
2. 40~45

3.

1. A 30
2. A A 90 ( B
A )
3. 60
4.
()

()()
1

Romanowsky-type staining technique

1. Basophilia (deep blue) affinity for a basic dye methylene

blue
Basophilia() DNA
RNA()
2. Azurophilia (purple) affinity for azure dyes; this is typical
of lysosomes.
Azurophilia()(azure dyes)

3. Eosinophilia (pink) affinity fro the acidic dye eosin;

Particular feature of hemoglobin which fills the cytoplasm of
erythrocytes.
Eosinophilia()(eosin)

4. Neutrophilia (salmon pink/lilac) affinity for a dye once

erroneously believed to be of neutral pH; characteristic of the
specific cytoplasmic granules of neutrophil leukocytes (leucocytes).
Neutrophilia() PH

4Erythrocytes (red blood cells)

5Functions of erythrocytes

Erythrocytes simply consist of an outer plasma membrane enclosing

hemoglobin and the limited number of enzymes necessary for
2

maintenance of the cell.

Oxygen and carbon dioxide transport.

Lifespan of an erythrocyte averages 120 days. 120

()

6Transmission electron micrograph showing

erythrocytes within a capillary

7Reticulocytes (immature red blood cells) in blood

RNA 24~48

ATP ()

8White blood cells (leukocytes, leucocytes)

Five types of leucocytes are normally present in the circulation.

These are traditionally divided into two main groups based on their
nuclear shape and cytoplasmic granules:

Granulocytes
1. Neutrophils
2. Eosinophils
3. Basophils
3

Mononuclear leucocytes
1. Lymphocytes
2. Monocytes

(azurophilc granules)

(gelatinase granules)

9Granulocytes

Granulocytes are so named for their prominent cytoplasmic secretory

granules.
Each of the three different types of granulocyte has type-specific
granules, the names neutrophil, eosinophil and basophil being derived
from the staining characteristics of these specific granules.
(neutrophil)
(eosinophil)(basophil)
The term polymorphonuclear leucocyte or polymorph as a synonym for
the term granulocyte.
(polymorphonuclear
leucocyte polymorph)

10Mononuclear leucocytes (agranulocytes)

()

Lymphocytes and monocytes, classified as mononuclear leucocytes,

have non-lobulated nuclei.

11Role of leucocytes as components of the body

defense systems

Acute inflammation (Neutrophils)

()(monocytes)

Neutrophils and monocytes are highly phagocytic and engulf

microorganisms, cell debris and particulate matter in a non-specific
manner.

(microorganisms)(cell debris)(particulate matter)

12Neutrophil

BY

Neutrophils are the most common type of leukocyte in blood and

constitute 40-75% of circulating leukocytes.
(leukocyte) 40-75
Principal function is in the acute inflammatory response to tissue injury
where they secrete enzymes that degrade tissue components, ingest
4

and destroy damaged tissue and kill invading microorganisms, particular

bacteria.
(acute inflammatory)

(a)
(b) D X D (drumstick
chromosome) 3%
* drumstick chromosome barr body
drumstick chromosome
(c)

(d) B (bacteria) Ps (pseudopodia)

(pus)

P (primary
granules)
5
S (secondary granules)
*

13Eosinophil

Eosinophils account for 1-6% of leukocytes in circulating blood.

1-6%
Eosinophil (12-17 m in diameter) is larger than the neutrophil and is
easily recognized by its large specific granules, which stain bright red
with eosin.
12-17 (eosin)(large
specific granules)
Specific granules are membrane-bound and of uniform size and the
matrix contains a variety of hydrolytic enzymes including histaminase.
(specific granules)(hydrolytic
enzymes)(histaminase)
The crystalloid has cubic lattice structure and consists of an extremely
alkaline protein called the major basic protein, other basic proteins,
hydrolytic lysosomal enzymes and a peroxidase termed eosinophil
peroxidase.
(cubic lattice structure)
(extremely alkaline )(major basic protein)
(hydrolytic lysosomal enzymes)
(eosinophil peroxidase)
D99.D100
primary granules (azurophilic granules )
(myeloperoxidase)
(neutrophil defenses)

secondary granules (specific granules )

tertiary granules (gelatinase granules )

(a) S specific granules

(b)
6

14Basophils

Basophils are the least common leukocyte and constitute less than 1%
of leukocytes in circulating blood.
1%
The basophil (14-16 m in diameter) is intermediate in size between the
neutrophil and eosinophil.
14-16
It has a bilobed nucleus but this is usually obscured by numerous large,
densely basophilic specific granules.

(basophilic specific granules)(obscured)

D99.D100

*
tobuldine blue
(histamine)

(a)
(b) S specific granules
15Lymphocytes

Lymphocytes are the smallest cells in white blood cells. They are the
second most common leukocytes in circulating blood and make up 2050% of the differential white blood cell count.
20-50%
Lymphocytes are characterized by a round, densely stained nucleus and
a relatively small amount of pale basophilic, non-granular cytoplasm.
(densely stained)

(a)

(b)


3%
(plasma cells)
(natural killer cells)

16Monocytes

Monocytes are the largest of the white blood cells (up to 20 m in

diameter) and constitute from 2-10% of leukocytes in peripheral blood .
20 (peripheral blood) 2%
They are motile phagocytic cells.
(motile phagocytic cells)
They are the precursors of macrophage found in peripheral tissues and
organs.
(macrophage)
(precursors)
Monocytes are characterized by a large, eccentrically placed nucleus.
Nuclear shape in variable but there is often a deep indentation of the
nucleus.
(eccentrically placed)
(deep indentation)
Cytoplasm contains numerous small lysosomal granules and cytoplasmic
vacuoles.
(small lysosomal granules)
(cytoplasmic vacuoles)
D99D100

M mitochondrion
G golgi apparatus
17Platelets

Gr cytoplasmic
grnules
Platelets
(thrombocytes)
are small, non-nucleated cells formed in the
P pseudopodia

bone marrow from the cytoplasm of cells called megakaryocytes.

(2m)(megakaryocytes)

()
Their numbers in circulating blood range from 150,000 to 400,000/ml.

150,000 400,000
Essential to the normal process of hemostasis, forming plugs to occlude
sites of vascular damage and promote clot formation.
(plugs)
(thrombin)(fibrinogen)(fibrin)

()(platelet disorders)
(thrombocytopenia)

(thrombocytosis)(thrombosis)

(a)
(b) dense granule
vesicle
(: alpha granules, dense granules, lysosomes,
microperoxisomes)

18 Bone marrow (PPT )

(red marrow)

()

(marrow stromal cells)

19Bone marrow smear, Giemsa stain

10

20Bone marrow, HE
stain

Aadipocytes Eendothelial cell Ssinusoid

Chemopoietic cords ()

()

21Different series of blood cell formation

11

()

(multipotential/pluripotential
stem cell)(unipotential stem cell)
Myeloblast

(~)Differentiation from stem cells to the

mature form of blood cells (
)

22Red cell formation (erythropoiesis)

(early normoblast)
(basophilic erythroblast)
(late normoblast)

12


()

23Granulocyte formation (granulopoiesis)

(myeloblast)(promyelocytes)
;

(stab cells)(band forms)

24Lymphocyte formation

(lymphoblast)
(prolymphocyte)

25Monocyte formation

-(CFU-GM)
(colony stimulating factors CSFs)

13

M1
M2
M3
M4

neutrophil myelocyte
metamyelocyte
stab cell
mature neutrophil

26Megakaryocyte and platelet formation

( 30~100m)

(megakaryoblast)

:
1.

(perinuclear zone N):

14

2.

(intermediate zone I):

(demarcation membrane systemDMS)

3.

(outer marginal zone): DMS

( )

(vesicle)
(DMS)
()

27Mature cell types in circulating blood

15

Chapter
4Connective tissue
DMS

1Introduction

Megakaryocyte
Connective tissue
provides structural and metabolic support for
plugs

other tissues and organs through the body.

clot
()

fibrinogen
Connective
tissues mediate the exchange of nutrients, metabolites,
fibrin

and waste
products between tissues and the circulatory system.
throbin

Connective tissues generally contain blood and lymphatic vessels.

2Components of supporting/connective tissue

Cells of connective tissue

Extracellular matrix(ECM)
consists of ground substance and fibers

3The cells of supporting/connective tissue

A dominant common function of these cells is synthesis and

maintenance of extracellular matrix material.

16

Fibroblast
responsible for secreting the extracellular matrix in most tissue
Ex
Chondrocytes and osteocytes
responsible for secreting the extracellular matrix in cartilage and
bone respectively.
chief cell ()

Adipocytes
for the storage and metabolism of fat.
5~10 ( 100 m)
Cells with defense and immune functions .
Ex : Mast cells, tissue macrophages, all types of white blood cells
and antibody-secreting plasma cells.

Tissue macrophages ( )

17

sub mucosa()
Fibroblast Collagen
Collagen

4The fibers of supporting/connective tissue

1. collagen (
reticulin)
2. elastin
Collagen
1. Type I collagen

2. Type II collagen
3. Type III collagen

4. Type IV collagen

5. Type VII collagen

Type I collagen 64nm

Reticulin fibers ( ) hepatocytes ( )

Recticular fiber type collagen 4~5%
(ex )
type

18

Elastin
(elastic lamina)

E
T
F
M
ML ( )

E
L
Cap
Ma

19

(a) E
(b)
()
(c)

(d)(EP)

(Ehlers-Danlons
syndrome)

Ground substance
/

Ground substance derived its

name from being an amorphous
transparent material which has
the properties of a semi-fluid gel.

Tissue fluid is loosely associated with ground substance, forming the

medium for passage of molecules.
(
p.69)
Ground substance consists of a mixture of long, unbranched
polysaccharide chains of seven different types of glycosaminoglycans

20


P.S.(glycosaminoglycans,GAGs)
(mucopolysaccharides)GAGs ()( p.69)

1. (hyaluronan) GAG
GAG GAG
(proteoglycans)
(mucoproteins) preteoglycans (90~95%)

2. GAG
( Na+)
(extracellular fluid)

GAGs

(proteoglycans)
21


fibrillin fibronectin
lamina,entactin tenascin
Ofibrillin
Fibronectin
laminin

Entactin laminin
Tenascin

6Basement membrane

Basement membrane are sheet-like arrangement of extracellular matrix

proteins which act as an interface between the support tissue and
parenchymal cells.
(
)
Basement membranes are associated with epithelial and muscle cells,
as well as forming a limiting membrane around the central nervous
system.

The term derives from the fact that the first basement to be recognized
were those lying beneath the base of surface epithelia.

External lamina()

()

(1)heparin sulfate
(2)Type IV collagen
(3)fibronectin (structural glycoprotein)

(cell adhesion, =integrins )

(4)laminin(non-filamentous proteins)

(5)entactin laminin Type IV collagen

Ps1. proteoglycan glycoprotein (
)()
Ps2. (glycoprotein)(fibronectin)
(fibrillin)
Ps3. (non-filamentous proteins)
lamininentactintenascin tenascin

22

 a PAS (Periodic Acid-Schiff stain)

/

b (renal tubules)(methenamine silver)

M mucus BM basement membrane

23

b a b
1. (lamina lucida)
2. (lamina densa) basal lamina
3. (lamina fibroreticularis)
basal lamina (basement membrane)
()

BM basement membrane
C collagen
E elastin
F fibroblast
Fc fibroblast cytoplasmic process
Ep epithelial
cell
Fibroblasts

IS intercellular space
IN basal invagination
LL lamina lucida
LD lamina densa
LF lamina fibroreticularis

()

ClCollagen fibers in longitudinal section

Primitive Mesenchyme
CtCollagen
fibers
transverse section

H&E in

ERRough
endoplasmic reticulum

mesenchymal
cellstem
cell
GGolgi
apparatus

(stellate)(fusiform)

loose connective tissue

24

Mature Fibroblasts
H&E stain

Collagenous Tissue

Loose connective tissue (areolar connective tissue)

a
b

aerolar connective tissue H&E

c trichrome

Dense connective tissue

25

e (!!
~)
Active FibroblastsH&E stain

(healing wound)(fibroblasts)
(myofibroblasts)

RER
scar tissue

1. PAS

2. HE Hematoxyline
Eosin
7Adipose Tissue
BY

Source

(primitive mesenchyme)
3. Massons trichrome
(lipoblasts)

Three main source

(1)Dietary fat circulating in bloodstream as chylomicrons.

(2)Triglycerides synthesized in the liver and transported in blood.

(3)Triglycerides synthesized from glucose within adipocytes.

26

Two main types of adipose tissue

(1) White Adipose Tissue
20% 25%
(energy store) (thermal
isolator) (cushion
against mechanical shock)
leptin
(2) Brown Adipose Tissue
(neonatal humans)
(hibernating animals)
()(adrenal gland
and great vessels in retroperitoneal fat)

(sympathetic nervous system)

White adipose tissue

(b)
standard
wax-embedded tissue preparation
(!)
N

(c)

D L

M
CCapillary
CoCollagen Fibrils
DSmall Lipid Droplet
FFibroblast Cytoplasmic Process
LLarge Lipid Droplet
Fibroadipose tissue
MMitochondria
NNucleus

Fibroadipose tissue
PCytoplasm
ofsupporting
Adipocyte
(collagenous
tissue) A
27

(lining)

Brown adipose tissue

(a,b)
S(Septum)

(a)

(b)
M

8 Defense Cells of Supporting

Tissue

1.

/ Fixed/intrinsic defense cell

MacrophageHistiocyteMast cell

2.

:(monocytes)

/Wandering/extrinsic defense cell

leucocytes

9Reticuloendothelial System
1. bone marrow
28

liverspleenlymph nodesthymus
2.
(1)
(2)
(3)
endothelial cell
3.
reticulum cell

Mast cell
(exocytosis)

histaminevascular activator

toluidine mast cell

granulemetachromasia
basophils
(1)heterochromatin
(2)glycogen
particles
a
metachromatic granules

b
G

Mi
GMast Cell Granule
MMast Cell
MiMitichondrion
Leukocytes
in loose supporting tissue

29

EoEosinophil
ErErythrocytes
FFibroblast
MaMast Cell
LLysosome

LyLymphocyte
McMacrophage
NNeutrophil
PcPlasma Cell
ppt !!

F
Er
N N
Eo

Ma
Ly
Pc

Mc

Macrophage
antigen
antigen presenting cell
antibody complement
opsonin
opsonisation
lymphokines

(cytokines)
T

30

 (EM)

L
R

Pp
Amoeba movement
Phagocytosis
LLysosome
rERRough Endoplasmic Reticulum
PpPseudopodia

PLatex particle
MMitochondrion
RResidual body

31