HUMAN HISTOLOGY (LABORATORY)

MODULE 5: CARDIOVASCULAR SYSTEM ANGELES, AJM.

REVIEW: BRIEF INTRODUCTION TO CARDIOVASCULAR

SYSTEM

The wall of typical artery contains three (3) concentric layer

tunics:

› TUNICA INTIMA – innermost layer facing the lumen

The cardiovascular system consists of the: › TUNICA MEDIA – middle layer
1. HEART and; › TUNICA ADVENTITIA – outermost layer

2. Closed system of BLOOD VESSELS that carry blood

which include:  Compared to arteries, veins are more numerous in the
body, have thinner walls with large diameter.
Major arteries ⎯ Aorta
Arterioles ⎯ Smallest kind of arteries  The walls of the veins also exhibit three (3) layers or
tunics. However, the muscular layer (tunica media) is
Capillaries ⎯ Smallest blood vessels
much thinner and less prominent.
Venules ⎯ Gradually increase in size to
become veins  Some muscular arteries exhibit two (2) thin wavy
Veins bands of elastic fibers called Internal Elastic Lamina
(IEL) and the External Elastic Lamina (EEL)

HEART
THREE LAYERS OF THE HEART
Quite easy to distinguish because of their clear differences in
thickness and in appearance

The walls of all four heart chambers consist of three major

layers: Section of right ventricle and the lower portion of pulmonary trunk

› The internal – endocardium EPICARDIUM

› The middle – myocardium
› The external – epicardium

› Outermost layer
› Staining properties: Clearly pale or light
staining

Composition:
A. Yellow line – Simple squamous mesothelium
→ Has single layer of squamous epithelium,
since it lines or surrounds the heart externally,
we refer to epithelium as mesothelium
B. Blue line – Underlying subepicardial layer of connective
tissue
→ This subepicardial layer contains numerous
coronary blood vessels, nerves, and adipose
tissue
→ Blue arrow: Coronary artery, which is a kind of
large blood vessel in the heart
→ Adipose tissue (fat) is easy to spot since it
appears white with lots of spaces inside each
adipocyte

MYOCARDIUM External epicardium with adipose showing thick myocardium which is

dark staining.

 Endocardium is not found in this specific tissue slide

because myocardium is too thick

LAYERS OF THE VASCULAR WALL

› Thickest layer – it would take a few fields to view

the entire myocardium.
› Sometimes, it is too thick that endocardium is cut
off or not even found in the tissue slide
› In all chambers of the heart, myocardium is the
thickest in the left ventricle
› Staining properties: stains darkest

Composition: TUNICA ADVENTITIA

A. Cardiac muscle fibers – branching cells are seen with
central nuclei
→ Green arrow: Intercalated disc
→ The myocardium in both atrium and ventricle
consists of cardiac muscle fibers

ENDOCARDIUM

Second image – Section of artery showing the vasa

› Innermost layer
› Located on the other end or
opposite of epicardium
› Endocardium lacks adipose tissue
unlike epicardium
Composition:
A. Simple squamous endothelium
B. Thin subendothelial connective tissue
Deeper to or below the endocardium, closer to
myocardium is the Subendocardial connective tissue,
wherein small blood vessels and Purkinje fibers are
found.
└ We usually use special stain to identify and
differentiate Purkinje fiver from regular cardiac
muscle fiber
└ Stain used in image: Mallory Azan Stain
vasorum because this is aorta (large artery), specifically
elastic artery
→ The red double lined arrow indicates the tunica
adventitia of both small artery and a small vein
Third image – A vein section also showing vasa vasorum
because it is a large vein
› Outermost layer of the blood vessel wall
› Composition: Longitudinally oriented collagen type
I fibers and elastic connective tissue fibers
› Consists primarily of collagen type I fibers
Tunica Adventitia of large arteries and veins contains:
⎯ A system of vessels that supplies
blood to the vascular walls
themselves
⎯ More extensive and needed in the
VASA walls of veins than arteries
VASORUM ⎯ Small blood vessels found in
tunica adventitia
⎯ The blood vessels of the blood
vessel

⎯ Network of autonomic nerves

NERVI
VASORUM
 TUNICA MEDIA
› Blue arrows – media of small artery and vein of
aorta
› First image – the darker stain of the media is due to
the elastic fiber present
› Second image – a large vein thin tunica media is
pointed with blue arrow
› Middle layer of the blood vessel wall
› Composition: Primarily circular smooth muscle
(SM) fibers and variable amounts of elastic fibers
└ A loose network of elastic fibers is
interspersed among the smooth muscle
cells
TUNICA INTIMA
› First image – The lighter staining later on top of
tunica media (yellow arrow) is the tunica intima,
although IEL is not well defined in the photo, it
should be around this part
› Second image – we can observe the thin
endothelium that lines the lumen of a small artery
and vein as well as the additional Internal Elastic
Lamina (IEL) in the artery
› Innermost layer that faces the lumen
› Composition: Simple squamous endothelium and
a thin underlying layer of a subendothelial layer of
connective tissue
The walls of some muscular arteries also exhibit TWO (2) THIN
BANDS OF ELASTIC FIBERS
ARTERY
EXTERNAL ELASTIC LAMINA (EEL)
This is a section of medium sized artery
 Light green arrow – indicates External Elastic
Lamina (EEL)
› It is located more externally
› Found at the periphery or at the border of the
muscular tunica media and is primarily seen in
large muscular arteries
› External Elastic Lamina (EEL) is present between
the media and adventitia of muscular arteries
› Described as a layer of elastin that separates the
Tunica Media from the collagenous Tunica
adventitia
INTERNAL ELASTIC LAMINA (IEL)
This is a section of medium sized artery
 Green arrow – indicates Internal Elastic Lamina
(IEL)
 Sometimes IEL is not seen in smaller arteries
› Found closer to the intima of blood vessel
› Found between the Tunica Intima and the Tunica
media
› Represents the most external layer of tunica
intima
› Layers of elastin sheets that contain numerous
openings or fenestrations
› Allow the rapid diffusion of nutritive substances
through the lamina to reach the cells deep within
the vessel walls
ARTERY VS VEIN
AORTA (ELASTIC ARTERY)
Section of aorta showing the:
Tunica intima – yellow arrow
Tunica media – blue arrow (thickest)
Tunica adventitia – red arrow
› The simple squamous endothelium and the sub-
endothelial connective tissue in tunica intima
remain unstained
› The first visible elastic membrane is the Internal
Elastic Lamina (IEL)
└ Since the aorta is elastic artery, its tunica
media is mainly composed of elastic fibers
than smooth muscle
└ Elastic fibers stain darker
└ Fine elastic fibers and smooth muscle
fiber are usually lighter stained or even
colorless
 › Tunica adventitia is also less stained and ARTERY VEIN
appears as a narrow, peripheral zone of ⎯ Innermost layer – tunica ⎯ The wall of the vein also
connective tissue in aorta intima stained dark contains tunica intima,
› The vaso vasorum is present consisting of └ Because of the thick media, and adventitia.
venules and arterioles Internal Elastic However, they are not
Lamina (IEL) which as thick as artery
› In such large blood vessels as the aorta and
is more prominent in ⎯ Tunica media – much
pulmonary arteries, the tunica media occupies most
arteries thinner and less
of the vessel wall
⎯ (Thickest) middle layer – prominent in the veins
› Whereas tunica adventitia is reduced to a ⎯ The smooth muscles
tunica media
proportionately smaller area intermix with connective
└ Contains several
layers of smooth tissue fibers
VEIN ⎯ Tunica adventitia –
arranged in a
circular pattern and thickest and best
thin dark strands of developed layer
elastic fibers ⎯ Surrounding both
⎯ Tunica adventitia – less vessels are the capillary,
conspicuous External arteriole, venule, and
Elastic Lamina (EEL) on cells of adipose tissue.
the periphery of tunica
media surrounding the
artery
⎯ Contains both light
staining collagen fibers
and dark staining elastic
Section of aorta showing the: fibers

Tunica intima – yellow arrow

Tunica media – blue arrow (thickest)
Tunica adventitia – red arrow

› Tunica intima of a vein consist of an endothelium

and supported by a small amount of sub-
endothelial connective tissue
› Large veins may exhibit an Internal Elastic Lamina
(IEL) that is not as well developed as in the Left = artery Bottom = vein
arteries  Prominent Internal  Even without
› Tunica media is thinner compared aorta endothelium lamina viewing the entire
(IEL) and; slide, just observe
› Smooth muscle fiber exhibit circular orientation
 Thick tunica media the layers and look
› In other large veins, the tunica media may be at the lumen
extremely thin and even more compact Right = vein  Tunica media is
› The thicker tunica adventitia is composed of  Larger lumen thinner
smooth muscle fiber in longitudinal orientation  Overall thinner  Lumen is wider and
 The fibers are segregated into bundles layers larger
and are seen mainly in cross-section  With adventitia as
surrounded by connective tissue of the its most developed Above = artery
tunica adventitia layer  Has more dark
 An arteriole, some venules, and capillary in staining elastic fiber
longitudinal section of vasa vasorum are
also visible in tunica adventitia This could be a muscular artery because the External Elastic
Lamina (EEL) is quite prominent.

PERIPHERAL BLOOD

OVERVIEW OF BLOOD

⎯ Blood is a fluid connective tissue that circulates

Illustration of muscular artery and vein in transverse plane
└ Elastic fiber = stained black
└ Collagen fiber = stained light yellow or yellowish
orange
Notice that wall of the artery is much thicker than the vein.
through the cardiovascular system
⎯ It is a specialized connective tissue consisting of cells
and extracellular material called plasma
 Like the other connective tissue, blood consist
of formed elements and an extracellular
component
⎯ Total blood volume in average adult: 6 liters
 7 - 8% of a total body weight
⎯ The heart's pumping action propel blood through
cardiovascular system to the body tissues
FUNCTIONS OF BLOOD
1. Nutrient and oxygen delivery to cells
 Directly or indirectly
2. Waste and CO2 transport away from cells
3. Delivery of hormones and other regulatory
substances to and from cells and tissues
 4. Acts as buffer to maintain homeostasis
5. Participate in immune responses
 coagulation and thermoregulation
 Transport of humeral agents and cells of
immune system that protect the body from
pathogenic agents, foreign proteins, and
transformed cells such as cancer cells
HOW IS BLOOD BEING PRODUCED?
 Blood cells have a limited lifespan. Hence, they wear
out and need to be continuously replaced in the
body
 HEMOPOIESIS – process of blood cell production
 This occurs in different organs of the body
depending on the stage of development of the
individual
 In developing embryo, hemopoiesis initially occurs in
the yolk sac and later in liver, spleen, lymph nodes,
and bone marrow
⎯ After birth, hemopoiesis continues almost
1. exclusively in the red marrow of different
bones
⎯ In the newborn, all bone marrow is red and
2.
functions in hemopoiesis
⎯ The red bone marrow is a highly cellular
structure and consists of hemopoietic stem
cells and the precursor of different blood
cells
3.
 Red marrow also contains a loose
arrangement of fine reticular
fibers that form an intricate network
⎯ As the individual ages and becomes an
adult, the red marrow is found primarily in
the flat bones of the skull, sternum, ribs,
vertebrae and pelvic bones
 The remaining bones or the long
4. bonds in the limbs of the body
gradually accumulate fat and their
marrow becomes yellow
 Consequently, they lose the
hemopoietic functions
 In hemopoiesis, all blood cells originate from
PLURIPOTENTIAL HEMOPOIETIC STEM CELL
 A common stem cell in the red bone marrow
that is self-renewing because it can produce
all blood cell types
 It produces two major cells lineages that
form the;
1. Myeloid stem cells
2. Lymphoid stem cells
 Before maturation and release into the bloodstream,
the stem cells from each lineage undergo numerous
divisions and intermediate stages of
differentiation
 In postnatal life, under normal circumstances, the
formation of cellular elements of the blood occurs in
the bone marrow in various bones
 Most of the proteins in plasma are made by the liver
 Plasma proteins → produced by liver
DEVELOPMENT OF PLURIPOTENTIAL HEMOPOIETIC
STEM CELL
⎯ Some lymphoid cells remain in the bone
1. marrow → proliferate → mature → become
B lymphocyte
⎯ While others leave bone marrow and migrate
via bloodstream → to lymph nodes and →
2. spleen → where they proliferate →
differentiate → to B lymphocytes after they
colonize peripheral lymphoid tissues
⎯ Other undifferentiated lymphoid cells
migrate to thymus gland, where they
proliferate and differentiate to
immunocompetent T lymphocytes.
3.
⎯ Afterwards T lymphocytes, would enter the
blood stream and migrate to reside in the
connective tissue and specific regions of
peripheral lymphoid organs of the body
⎯ Both B and T lymphocytes reside in
numerous peripheral lymphoid tissues,
4. lymph nodes, and spleen
 They initiate immune responses
when exposed to antigens
WHAT HAPPENS TO SENESCENT CELLS (OLD CELLS)?
 The pluripotential hemopoietic stem cells
continually divide and differentiate to produce new
progeny of cells because all blood cells have a limited
lifespan.
 When blood cells become worn out and die, they are
destroyed by macrophages in different lymphoid
organs such as the spleen
 FOUR MAJOR ELEMENTS OF BLOOD
› Erythrocytes
Red blood › Transport oxygen from the lungs
cells to the peripheral tissues
› Leukocytes
› Have a defensive role, destroying
infecting organisms, such as
White blood
cells bacteria and viruses, as well as
assisting in the removal of dead or
damaged tissues
› Thrombocytes
› Are the first line of defense
against damage to blood vessels,
Platelets adhering to defects and
participating in the blood clotting
system (coagulation)
› Is the proteinaceous solution in
which the above-mentioned cells
circulate, and carries nutrients,
Plasma metabolites, antibodies, hormones,
proteins of the blood clotting
system and other molecules
throughout the body.
HOW DO WE HISTOLOGICALLY EXAMINE BLOOD?
 Blood cells can be studied histologically in smears
prepared by spreading a drop of blood in a thin layer
on a microscope slide.
BLOOD SMEAR PREPARATION
Image: two blood smears
⎯ Examination of blood cells requires special
preparation and staining
 This method differs from the usual preparation
seen in histology laboratory, in which the
specimen is not embedded in paraffin and
section
 Rather a drop of blood is placed directly on
a slide and spread thinly over the surface,
and then pull with the edge of another slide,
it will then produce a monolayer of cells. The
preparation is then air dried and stained.
→ Area of interest = monolayer of cells
(thin part of smear)
 Another difference in the preparation of blood
smear is that special mixture of dyes is used
to stain the blood instead of using traditional
hematoxylin and eosin stain
⎯ For laboratory purposes, samples of blood are often
drawn from a vein
⎯ Blood smears are routinely stained with a mixture of
acidic (eosin) and basic (methylene blue) dyes =
AZURES
 These mixtures may also contain dyes called
azured that are more useful in staining
cytoplasmic granules
 Cytoplasmic granules are helpful in
characterization of different types of RBC
PREPARATION OF BLOOD SMEAR
› Prick a finger and collect a small amount
STEP 1
of blood using a micro pipette
STEP 2 › Place a drop of blood on a slide
› Using a second glass slide, pull the drop
of blood across the first slide surface,
leaving a thin layer of blood on the slide
STEP 3
› After the blood dries, apply stain briefly
and then rinse and place coverslip on
top
› Viewed under microscope revealing
STEP 4 components of formed elements (blood
cells)
RED BLOOD CELLS
(ERYTHROCYTES)
⎯ Mainly responsible for O2 transport
⎯ Appear as rounded, bright pink stained cells
(acidophilia)
 Bright pink color is due to the content of
oxygen carrying hemoglobin which binds
the acidic eosin dye used in staining
⎯ 6.5 – 8.5 um in diameter
⎯ Size decreases slightly with age
⎯ Biconcave shape, pale center, darker periphery
 Biconcave shape maximizes their surface
area or volume ratio. Hence, maximizing
oxygen exchange
⎯ Erythrocyte are also quite flexible and can easily
bend to pass through the small capillaries, which
stock up in loose aggregates (stock coins) which
called Rouleaux
⎯ Lack nuclei (during hemopoiesis)
 Nuclei is lost during formation
⎯ Metabolically active despite lack of organelles
 There are no discernible organelles as they
are lost during differentiation
⎯ They are metabolically active and derive energy by
anaerobic anabolism of glucose and through
ATP generation hexose monophosphate shunt
 ⎯ Main cytoplasmic constituent: Hemoglobin WHITE BLOOD CELLS
 A protein complex which results in its (LEUKOCYTES)
characteristics acidophilic staining
property
⎯ RBC membrane: Actrin/spectrin
 Containing cytoskeletal meshwork
 Cell membrane is braced by actrin or
spectrin containing cytoskeletal meshwork
which is largely responsible for maintaining
the distinctive biconcave shape
⎯ Lifespan: 100 - 120 days in the circulation
 Mature red cell is unable to synthesize new
enzymes to replace those lost during
⎯ Leave the blood and migrate to the tissues where
normal metabolic processes
they become functional and perform various
 The diminishing efficiency of ion activities related to immunity
pumping mechanisms is probably the
⎯ Divided into two (2) major groups:
main factor in red cell aging
 Granulocytes and Agranulocytes
⎯ The cells becoming less deformable until it is
− They are classified based on the
unable to negotiate microcirculations and
density of their cytoplasmic
further removed by phagocytosis
granules
⎯ Sites of RBC sequestration:
⎯ In circulation, their shape is spherical
1. Spleen
⎯ In tissues, they can be ameboid in shape
2. Liver
3. Bone marrow
GRANULOCYTES AGRANULOCYTES
⎯ All dispose of aged and defective red cells
⎯ Spleen: most active site of RBC sequestration › They possess two major › Lack specific
types of abundant granules but do
cytoplasmic granules: contain some
› Abnormal arrangement of
internal RBC cytoskeleton, › Azurophilic – lysosomes azurophilic
granules
causes deformation due to › Specific granules bind
(lysosomes)
ankyrin/spectrin abnormality neutral basic or acidic
stains and they have › Spherical nucleus
Hereditary › Abnormal round convex blood
Spherocytosis specific functions also or intended but not
cell, brittle and less deformable
lobulated
› Cannot freely pass-through › Characterized by
polymorphic nuclei with › Monocytes and
microcirculation, they are trapped
two (2) or more distinct lymphocytes
and destroyed
lobes
› Caused by iron deficiency, less › Neutrophils, Eosinophils,
Anemia hemoglobin than normal, pale Basophils
staining (hypochromic), smaller › All granulocytes are
size, larger central pallor, thinner terminally differentiated
periphery cells with a life span of
› Most common blood disorder only few days.
› Inadequate hemoglobin supply
causes weakness, pallor and
sometimes breathlessness

› Result from either impaired red

cell formation or excessive red
cell destruction
› Most common cause:
Deficiency of iron
 Iron is essential in the
formation of GRANULOCYTES
hemoglobin
Other › Other possible causes: NEUTROPHILS
mutations such as patients with
sickle cell anemia

Sickle cell anemia – due to abnormal

hemoglobin causing RBC to be sickle in
shape which may not survive for long
and can also block blood vessels
⎯ Most numerous WBCs (50-70% of circulating
leukocytes)
⎯ Most common granulocytes
⎯ Measure 10 - 12 um in diameter in blood smear
⎯ Larger than erythrocytes
⎯ Polymorphonuclear neutrophils / polymorphs: 2
– 4 lobes
⎯ Although named for their lack of characteristic
cytoplasmic staining, they are readily identified for
multimodal nucleus.
 Thus, they are also called
Polymorphonuclear neutrophils or
polymorphs
 ⎯ Neutrophils possess 2 - 4 lobes of nuclear material ⎯ In blood smears, eosinophils is about the same
joined by thinner nuclear strands size as neutrophil or slightly larger but with a
⎯ Life span of 1-4 days in connective tissue before characteristic bilobed nucleus
dying by apoptosis ⎯ Main identifying characteristic: Abundance of
⎯ They are short lived cells with half-life of 6 - 8 large acidophilic specific granules typically
hours in blood staining pink or red
⎯ Specific granules are oval in shape with
Neutrophils contain three types of granules: flattened crystalloid cores – has major basic
proteins (MBPs) and eosinophilic peroxidase
› Azurophilic (primary) -
myeloperoxidase, acid › An arginine rich factor
hydrolases, defensins, MBP accounts for granules acidophilic
*cathelicidin and constitutes up to 50% of
 Lysosomes of neutrophil total granule protein
 Larger and less Eosinophilic › Act to kill parasitic worms or
numerous than specific peroxidase helminths
granule ⎯ Hence, eosinophils also modulate inflammatory
 Arise early in responses by releasing chemokines, cytokines,
granulopoiesis and lipid mediators triggered by allergies oe
hypersensitivity reactions (key words)
 Occur in all granulocytes
⎯ Number of circulating eosinophils would increase
as well as in monocytes during helminthic infections or allergic
and lymphocytes reaction
PRIMARY ⎯ Abundant in connective tissue of intestinal lining
− Helps generate
and sites of chronic inflammation such as lung
highly reactive
tissues (asthmatic patients)
bactericidal
Myeloperoxidase
hypochlorite and ⎯ It also removes antigen-antibody complexes
from interstitial fluid by phagocytosis
chloramines
BASOPHILS
− Cationic proteins
− Function
analogously to
antibodies and
Defensins antimicrobial
peptide called
cathelicidin to
kill pathogens ⎯ Measure 12 - 15 um in diameter
⎯ Basophils are the least numerous of the WBCs,
› Specific (secondary) - accounting for less than 0.5% of total leukocytes.
collagenase, phospholipase, ⎯ Difficult to find in normal blood smear
complement activators, ⎯ Has two (2) irregular lobes but large specific
lysozymes, lactoferrins granules overline the nucleus, usually obscure
its shape which stain purple
SECONDARY  Smallest granules
 Twice as numerous as  Similar with eosinophil with two (2) lobes,
azurophilic granules but in basophil, specific granules are
larger and mask nucleus. Hence, two
 Contain various enzymes
lobes is not that visible.
⎯ The specific granules typically stain purple with
› Tertiary – phosphatases, basic dye of blood smear stains and are fewer,
metalloproteinases larger, and more irregularly shaped than the
granules of other granulocytes
− Enzymes that ⎯ Strong basophilia due to heparin
remove a ⎯ Granules also contain histamine and other
phosphate mediators of inflammation
group from a  Including platelet activating factor,
Phosphatase
substrate eosinophil chemotactic factor, and;
− Sometimes called  Enzyme phospholipase A - catalyzes an
TERTIARY phosphosome initial step in produced lipid-derived
proinflammatory factors called leukotrienes
− Include ⎯ By migrating in connective tissue, basophil appear
gelatinases and to supplement the functions of mast cells
collagenases ⎯ Both basophil and mast cells have metachromatic
Metalloproteinases
− Facilitate granules which contains heparin and histamine.
migration of  They have surface receptors for
neutrophil through immunoglobulin E and secrete granular
connective tissue components in response to certain antigens
and allergens
EOSINOPHILS

AGRANULOCYTES

LYMPHOCYTES

⎯ Far less numerous than neutrophils, only 1%-4%

of leukocytes
 PLATELETS
(THROMBOCYTES)

⎯ Most common agranulocytes

⎯ 30% of total blood leukocytes
⎯ Lymphocytes are the main functional cells of the ⎯ Are small, membrane-bounded, anucleate
lymphatic or immune system cytoplasmic fragments derived from
⎯ Strong basophilia due to heparin megakaryocytes
⎯ Most lymphocytes found in blood or lymph ⎯ In platelet formation, small bits of cytoplasm are
represent recirculating immunocompetent cells separated from peripheral regions of
⎯ Cells that have developed the capacity to recognize megakaryocyte
and respond to foreign particles to antigens are ⎯ Platelets function in continuous surveillance of
in transit from one lymphatic tissue to another blood vessels, blood clot formation, and repair of
 In tissues associated with immune system, injured tissue
three (3) groups of lymphocytes can be ⎯ They are involved in several aspects of
identified according to size hemostasis (control of bleeding)
⎯ In blood smears, the mature lymphocytes ⎯ Platelets continuously survey the endothelial
approximate the size of an erythrocyte lining of blood vessels for gaps and breaks

› Have intensely staining, slightly Example:

indented spherical nucleus,
cytoplasm appears very thin pale Injured blood vessel wall → promote platelet adhesion on
Small blue rim surrounding the nucleus exposed connective tissue at damaged site → triggers
degranulation → release of serotonin, ADP, and
lymphocytes › No recognizable cytoplasmic
thromboxane A2
organelle other than occasionally
(rare) fine azurophilic granules
⎯ Often found as aggregates
Medium
lymphocytes
› Either activated lymphocytes
Large which possess surface receptors
lymphocytes that interact with specific antigen
or a natural killer lymphocytes

MONOCYTES

⎯ Monocytes are the precursors of the cells of the

mononuclear phagocytotic system
⎯ Largest of the WBCs in a blood smear
⎯ Average diameter of 18 um
⎯ They travel to bone marrow to body tissues
where they differentiate into the various
phagocytes of the mononuclear phagocytotic
system
⎯ We have connective tissue macrophages,
osteoclasts, alveolar macrophages, perisinusoidal
macrophages in the liver (Kupffer cells)
 But they are also present in lymph nodes,
spleen, and bone marrow
⎯ Life span in blood: 3 days
⎯ More indented nucleus than lymphocyte (horse-
shoe shaped, bean-shaped)
 The indentation is the site of the cell
center where the well-developed Golgi
apparatus and centrioles are located
⎯ Also contain Smooth ER, Rough ER, and small
mitochondria
⎯ Has small dense azurophilic granules
⎯ Typical lysosomal enzymes found in azurophilic
of neutrophils
 HUMAN HISTOLOGY LABORATORY
MODULE 6: DIGESTIVE SYSTEM
MODULE LEARNING OUTCOME
At the end of this module, students should be
able to:
• Identify and differentiate the distinct
histological features of the different
tissues under the Digestive System.
LECTURE CONTENT
• The mucosa is lined by non-keratinized
stratified squamous epithelium.
• In its submucosa, it usually contains
esophageal glands.
• It has an inner circular and outer longitudinal
muscle layer (skeletal muscle layer) and it is
covered by adventitia.
MIDLLE ESOPHAGUS

• Introduction
• Esophagus
• Stomach
• Small Intestine
• Large Intestine

ESOPHAGUS

• Its epithelium is also non-keratinized stratified

squamous epithelium.
• It has a submucosa that also contains
esophageal glands but its not appreciated
in this histologic picture or micrograph.
• It has an inner circular and outer longitudinal
muscle layer which are mixed skeletal and
smooth muscle layer.
• It is also covered by adventitia which is not
quite appreciated in this micrograph.
• The esophagus has several histologic layers
which are the following: mucosa,
submucosa, muscularis externa and LOWER ESOPHAGUS
adventitia or serosa depending on the
region of the esophagus.
• The mucosa of the esophagus has a non-
keratinized stratified squamous epithelium
and its submucosa would usually contain
the esophageal gland.
• There are several differences as mentioned
in the regions of the esophagus mainly the
upper, middle and the lower esophagus
particularly in its muscularis externa.

UPPER ESOPHAGUS

• Also lined by stratified squamous epithelium,

non-keratinizing.
• In this layer, it has a transition between a
stratified squamous into a simple columnar
epithelium of the stomach.
• The submucosa would also contain several
esophageal glands and the muscularis
layer or externa is exclusively smooth
muscle.
• In this region, it is covered or lined by serosa
rather than adventitia.

Page 1 Cortez &Hernandez

 • It has three arrangements of its muscularis
externa, an inner oblique, middle circular
STOMACH and an outer longitudinal smooth muscle.
CARDIA REGION; EGJ

BODY REGION

• As mentioned, the esophagus is lined by • The body is quite similar also with the fundic
stratified squamous epithelium and it will region.
transition into simple columnar epithelium of • In its mucosal layer as well as the usual cells,
the stomach which usually has gastric pits. the chief glands in the lamina propria.
• Mucus-secreting of cardiac glands are
usually located in the lamina propria.
• We usually don’t expect any glands in the PYLORIC REGION
submucosa layer but rather connective
tissue with large blood in the vessels as well
as lymphoid cells, macrophages and mast
cells which are not appreciated in this
micrograph.
• The muscle layers are usually inner oblique,
middle circular and outer longitudinal
smooth muscle are not well defined.
• The stomach in this region is also covered by
serosa.

FUNDIC REGION

• It is lined by simple columnar epithelium with

gastric pits.
• Its mucus-secreting cells of pyloric glands
are also located in the lamina propria.
• It is lined just like the rest of the region with
serosa.
• Its muscularis layer, which is not seen in the
picture, has the arrangements.

• Quite similar when we talk about Histology SMALL INTESTINE

with the body of the stomach.
• It is also lined by simple columnar epithelium
in its mucosa with noticeable gastric pits.
• It has parietal cells and chief cells of gastric
glands in its lamina propria.
• Just like the other regions of the stomach, no
glands are expected in the submucosa but
usually large blood vessels and lymph
vessels are seen as well as several cells such
as lymphoid cells and macrophages.
• It is also lined by serosa.

Page 2 Cortez &Hernandez

 • It has a bruner or duodenal glands in its
• Small intestine has a several regions: submucosa
• It has columnar absorptive goblet cells in
1. Duodenum
mucosal layer
- Expect submucosal layer

- Bruner or duodenal glands

JEJUNUM
2. Jejunum

3. Ileum

• Small intestine mucosa is lined by columnar

absorptive and goblet cells
• Other cells are also seen in the mucosal layer
such as enteroendocrine cells

MUSCULARIS EXTERNA

• Similar mucosal layer lining

• Submucosa layer usually have no glands
• Muscularis externa has an inner circular
muscle and outer longitudinal muscle that is
well defined

ILEUM

• Has two arrangement or layers:

1.Inner circular

2.Outer longitudinal smooth muscle

• Myenteric plexus aka Auerbach Plexus or

back nerve plexus

-Responsible for the muscular contraction

SUBMUCOSAL PLEXUSES

• Cells from enteric nervous system
REGIONS OF THE SMALL INTESTINE
DUODENUM
• Submucosa has a several Peyer patches or
lymphoid nodules
• It has the same lining as the rest of the
regions of the small intestine
• It also has a similar lining in its outer layer
which is the serosa

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 LARGE INTESTINE • Attached to the cecum
• It contains mucosa, submucosa and
muscularis externa
• Large intestine has a several regions:
• Lymphatic nodules are usually noticeable
1.Cecum

2.Colon
ANORECTAL JUNCTION
3.Rectum

4. Anal Canal

• The lining epithelium or the mucosa is lined

by goblet cells and columnar absorptive
cells
• It has numerous goblet cells compared to
the small intestine
• It has a submucosal layer which usually has
no glands in the sickle region as well as the
colon and the rectum
• It Muscularis Externa has an inner circular
smooth muscle and outer longitudinal
smooth muscle
• It layers forms three thin yakuli

• The transition from simple columnar

epithelium from the rectal area into
Stratified squamous epithelium in the anal
canal is seen.
• In the rectal area in the submucosa, from
the cecum to the rectal area, we would
usually not expect any glands in its
submucosa layer
• Expect sebaceous glands and venous
structures in anal canal

APPENDIX

Page 4 Cortez &Hernandez

 MTHumHis Lab M7
Urinary System Vascular Pole Entry point of the
glomerular capillaries
Kidney into the adrenal
corpuscle
Afferent arterial enters
and the efferent
arteriole exits the renal
corpuscle
Urinary Pole On the opposite end of
the vascular pole of the
renal corpuscle
Where the filtrate
produced by the
glomerulus lives in the
renal corpuscle
Renal Tubules The glomerular filtrate
that leaves the renal
corpuscle firsts enters
the renal tubule which
extends from the
glomerular capsule into
the collecting tubule
Has several distinct
histologic and functional
regions
Proximal Portion of the renal
This micrograph or Convoluted tubule that starts at the
histologic section Tubules renal corpuscle is highly
shows the major twisted
histologic features of a Located in the cortex
renal corpuscle but then descents into
the medulla to become
continuous with another
tubule, the loop of henle
Glomerulus Renal corpuscle Loop of Henle Consists of several
consists of a tuft of parts: a thick portion of
capillaries the proximal convoluted
Glomerular or Surrounding the tubule, a thin
Bowman glomerulus by a double descending and
Capsule layer of epithelial cells ascending segment,
Podocytes Inner or visceral layer of and a thick ascending
the capsule consists of portion called the distal
unique and highly convoluted tubule
specialized branching Distal Shorter and less
epithelial cells Convoluted convoluted than the
Outer or parietal layer of the glomerular Tubule proximal convoluted
capsule consists of simple squamous tubule
epithelium Ascends back into the
Macula Densa Near the corpuscle kidney cortex
Renal Initial segment of each From the distal
Corpuscle nephron convoluted tubule to
glomerular filtrate flows
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to the collecting tubule
which is not seen here
Collecting tubule and the collecting duct are
not part of the nephron
This micrograph or histologic section shows
the continuity of the renal corpuscle tubular
pole between the simple cuboidal
epithelium of the proximal convoluted
tubule and the simple squamous epithelium
of the capsule parietal cell
Ureter and Bladder
A diagram of the cross-section of the ureter
showing the characteristic pattern of
longitudinally folded mucosa
MTHumHis Lab M7
The urinary space or the bowman space
between the parietal layer and the
glomerulus would drain into the lumen of
the proximal convoluted tubules
The lumens of the proximal convoluted
tubules appear filled because of the long
microvilli of the brush border and
aggregates as well of small plasma
proteins that are bound to this structure
In contrast to the lumen, it appears empty
and lacks a brush border as well as routine
Proximal Has long microvilli of its
Convoluted brush border
Tubule
Distal Lacks brush borders
Convoluted
Tubule
This is surrounded by a thick muscularis
layer that moves the urine by regular waves
of peristalsis
The ureter’s mucosa consists of transitional
epithelium or urothelium and a wide lamina
propria
The transitional epithelium has several
layers
The outermost layer is characterized by
large cuboidal cells
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The intermediate layers are polyhedral in
shape whereas the basal layers or basal
cells are low columnar or cuboidal
MTHumHis Lab M7
This thick epithelial lining varies between
stratified columnar in some areas and
pseudostratified columnar elsewhere but it
becomes stratified squamous at the distal
end of the urethra

This is a micrograph for histologic section

of the bladder which shows the four
histologic layers of the bladder
In the neck of the bladder near the urethra,
the wall shows these four layers: mucosa
with urothelium and lamina propria, a thin
submucosa, an inner layer, middle layer, an
outer layer of smooth muscle,and the
adventitia
Bladder is empty The mucosa is
highly folded and
the urothelium has
bulbous umbrella
cells
Bladder is full The mucosa is
pulled smooth and
the urothelium is
thinner and the
umbrella cells are
flatter

Urethra

This micrograph shows the urethra

The urethra is a fibromuscular tube that
carries urine from the bladder to the
exterior of the body
Transverse section of the urethra showing
the mucosa that has large longitudinal folds
In the higher magnification, it shows the
unusual stratified columnar nature of the
urethral epithelium

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 MTHumHis Lab M7
pelvis to form the renal
papilla
Urinary System Papilla Where urine is
collected
Minor Calyx After collected, it will
be in the minor calyx
Smallest branches
The one who is directly
connected in the renal
pyramid
Then minor calyx will
join together to form
the major calyxes
Major Calyxes Usually situated in the
middle
When two or more
minor calyxes joined
The one that will
proceed to renal pelvis
Composed of the 2 kidneys, 2 ureters, while going through our
urinary bladder, and urethra hilum
Main function Production, storage, Then, the urine will
and expulsion or leave in the ureter
excretion of urine
Other functions Secretion of renin,
angiotensin, vitamin
B, and erythropoietin

Regulation and
endocrine

Kidney

This is a renal pyramid

Renal Capsule The thin layer of dense
irregular connective
tissue
Has cortex and medulla Cortex Outer part and darker
Cortex Outer part of an organ in color or dark-staining
Medulla Inner part of an organ Renal Consists of glomerulus
Renal Pyramids Embedded in the Corpuscles and Bowman’s capsule
kidneys Convoluted Proximal Convoluted
Base is facing the Tubules Tubules and Distal
cortex Convoluted Tubules
Apex extends Medullary Rays Straight tubules and
downwards to the renal collecting tubules

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 MTHumHis Lab M7
Medulla Inner part and lighter- JGA JG Cells
staining (Juxtaglomerula (Juxtaglomerular
Pyramid Base – Cortex; Apex – r Apparatus) – Cells) – modified
Papilla maintains the smooth muscle cells
Straight Tubules Collecting Ducts blood pressure to with cytoplasmic
Cortex ensure adequate granules
glomerular blood Macula Densa –
flow rate for the narrow, more columnar
filtration and cells that border the JG
reabsorption of cells
substances in the
glomerulus

Where we will see the glomerulus and

Bowmans capsule
Cortex: PCT & DCT
Glomerulus Tuft of capillaries
surrounded by a
double layer of
epithelial cells which is
the Bowman’s capsule
or glomerular capsule
Inner or Visceral Where we can find the
Layer of the podocytes
Bowman’s
Capsule
Cortex: Renal Corpuscle
Proximal Round to oval
Convoluted Simple cuboidal
Tubule epithelium
Many mitochondria –
granular and intensely
eosinophilic
Many microvilli –
prominent brush border
Epithelial Part Bowman’s Capsule –
Cells are larger and
simple squamous
more elongated than
epithelium
DCT
Bowman’s Space – Smaller uneven lumen
tiny space for the urine; (the hole of the tubule)
that’s where urine
secreted by the
glomerulus; also called
as urinary or capsular
space
Vascular Part Tuft of glomerular
capillaries from the
afferent arteriole
Where blood is filtered Distal Simple cuboidal
to be excreted later on, Convoluted epithelium
on the form of urine Tubule Less eosinophilic
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 MTHumHis Lab M7
Smaller diameter
Wider lumen
No brush border
Cells are smaller and
more closely packed
than PCT
Small and even lumen,
usually perfect circle
Medulla: Loop of Henle
Lesser in number in
Thin Segment Simple squamous
the renal cortex
epithelium
Thick Segment Simple cuboidal
epithelium
Medulla: Collecting Ducts
Collecting Ducts Simple cuboidal or
columnar epithelium
Less specialized than
PCT and DCT
Clear cytoplasm
Distinct cell borders
Different sizes:
smallest in cortex;
largest near the renal
pelvis
Principal Cells – pale-
staining, columnar cells
rich in ADH-regulated
saquaporins for water
Medulla absorption
Intercalated Cells –
fewer, darker cells with
projecting apical folds

Ureter

Collecting Ducts Straight portions of the

and Vasa Recta tubule
Loop of Henle Thick and thin segment

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 MTHumHis Lab M7
Has 3 major layers: mucosa, smooth Adventitia
muscle, adventitia Outer portion of the ureter
Mucosa Layer Blends with fibroelastic connective tissue
and adipose
Arterioles, venules, and small nerves

Urinary Bladder

Transitional Has the ability to

Epithelium stretch or change its
shape depending on
the pressure or the
volume of the filtrate Serves as a container or catch basin for the
that it receives from urine or the filtrate
the kidney Has a thick layer of transitional epithelium
Outermost layer: It has to cater to a large volume of liquid so
large cuboidal cells it has to have the ability to stretch or adjust
Intermediate layer: its size
polyhedral cells Its histology depends if it is extended or
Basal layer: low filled or empty or relaxed
columnar or Mucosa (Empty Bladder)
cuboidal Transitional Presence of mucosal folds
Wide Lamina Fibroelastic Epithelium Once the bladder is
Propria connective tissue (5-7 layers) distended, it becomes flat
Superficial cells: dome-
shaped, binucleate
Intermediate cells: round
Basal cells: more columnar
Lamina propria: wider than
the ureter’s; has to support
the transitional epithelium

Muscularis
Upper Ureter (2 Closer to the kidney
layers) Inner longitudinal
smooth muscle
Middle circular
smooth muscle Mucosa (Filled Bladder)
Lower Ureter (3 Closer to the Transitional Folds in the bladder wall
layers) bladder Epithelium disappear
Inner longitudinal (2-3 layers) Basement membrane is not
smooth muscle folded
Middle circular Contains urine already
smooth muscle Surface cells flatten
Outer longitudinal Intermediate cells are
smooth muscle pushed and pulled laterally

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 MTHumHis Lab M7
Resembles stratified Loose connective tissue with autonomic
squamous epithelium nerves and plexuses, blood vessels, and
lymphatics

Mucosa (low magnification)

Lamina Compose of connective
Propria tissue fibers Female Urethra
In the deeper layers, more
elastic fibers
Fibroblasts
Blood vessels, venule, and
arteriole
Muscularis (Detrusor Muscle)
Has 3 delineated layers which mean the 3
layers are distinguishable from each other
Inner longitudinal smooth muscle
Middle circular smooth muscle
Outer longitudinal smooth muscle
When it contracts, it is the feeling of the
urge to urinate

Serosa or Adventitia
Both cover the outer layer of the urinary
bladder but the difference is the location
Serosa Upper part of the bladder
Roof of the bladder
Adventitia Lower part of the bladder
Loose connective tissue
that covers the part of the
bladder with no serosa
Mucosa
Transitional Near the bladder
Epithelium
Stratified Columnar Intermediate zone
Non Keratinized Opening
Stratified Squamous
Lamina Propria
Muscularis
Inner longitudinal smooth muscle
Outer circular smooth muscle
Adventitia
Thin layer of connective tissue

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 MTHumHis Lab M7
Male Urethra

Male Urethra
Cavernous Spaces
Composed of cuboidal
Glands of Littre cells
Secretes mucous

Much longer than female urethra

More complicated histologically and
anatomically
Since the ducts for sperm transport
during ejaculation joins the urethra at
the prostate gland
Prostatic Urethra 3-4 cm long
Lined by urothelium
Membranous Lined by stratified
Urethra columnar and
pseudostratified
columnar
epithelium
Spongy Urethra 15 cm long
Lined by stratified
columnar,
pseudostratified
columnar, and
stratified squamous
distally

Female VS. Male Urethra

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