MLS 2206 - Human Histology College of Medical Laboratory Science - Central Philippine University

MODULE 1
INTRODUCTION
MLS 2206 | Human Histology

College of Medical Laboratory Science | Central Philippine University
MLS 2206 | HUMAN HISTOLOGY

Module 1 - Introduction
1.1 – Introduction to Histology

1.2 – The Cell

Module 1.1 – Histology and
Student Learning Outcomes
its Methods of Study
1.1.1. Meaning of Histology 1. Understand comprehensively the field

1.1.2. Meaning of Histopathology of histology
1.1.3. Meaning of Cytology 2. Relate logically the study of
1.1.4. Steps involved in the preparation of tissues histopathology to histology
1.1.4.1. Fixation
3. Define correctly the term cytology
1.1.4.2. Dehydration
4. Determine systematically the steps
1.1.4.3. Clearing
involved in the preparation of tissues
1.1.4.4. Infiltration
1.1.4.5. Embedding
for microscopic examination
1.1.4.6. Blocking 5. Apply meticulously the techniques
1.1.4.7. Trimming involved in microscopy
1.1.4.8. Cutting
1.1.4.9. Staining
1.1.4.10. Mounting
1.1.4.11. Labeling
1.1.5. Techniques in microscopy of the prepared
slides (to be discussed in the lab class)
Meaning of Histology
Histology
• comes from the Greek word “histo”
meaning “tissue” or “web”
• the study of the tissues of the body and

how these tissues are arranged to
constitute organs.
• Involves all aspect of tissue biology, with

the focus on how cell’s structure and
arrangement optimize functions specific to
each organ. MLS 2206 | HUMAN HISTOLOGY
EPITHELIAL CONNECTIVE MUSCLE NERVOUS
Four Basic Types of Tissues

Tissues are made up of 2
interacting components:
Cells
Extracellular matrix
• Highly organized molecules that
form complex structures
• Functions:
✓ Furnish mechanical support for
the cells
✓ Transport nutrients to the cells
✓ Carry away catabolites and
secretory products MLS 2206 | HUMAN HISTOLOGY
Meaning of Histopathology
Histopathology
• deals with the study of the basic disease
process, correlating the etiology of
disease with the course development of
anatomic and clinical changes brought
about by disease.
Cloudy swelling of the renal tubules. The outline of the

tubular lumen is blurred.

Meaning of Histopathology
Histopathologic Techniques
• Deals with the preparation of tissues for
microscopic examination.
• Involve different procedures that have been

adopted for the preparation of tissues for
microscopic investigation
AIM OF HISTOPATHOLOGICAL TECHNIQUE:

• produce microscopic preparations of tissues usually
stained, that represent as closely as possible their
structures.
Meaning of Cytology
Exfoliative Cytology
• branch of science that deals with the
study of cells that have been
desquamated from epithelial surfaces
originated from exfoliative cells coming
from the lining epithelium and mucosa of
different organs of the body.

Meaning of Cytology
Cytopathology
• study of abnormal cells as in cancer cells
Cytotechnique
• study of the methods of preparing the
cells for microscopic examination.

Preparation of Tissue for Study
TISSUE PROCESSING
• Tissues from the body taken for diagnosis of
disease processes must be processed in the
histology laboratory to produce microscopic slides
that are viewed under the microscope by
pathologists.
HISTOTECHNIQUES
• The techniques for processing the tissues, whether
biopsies, larger specimens removed at surgery, or
tissues from autopsy
• Histotechnologists.
SECTION CUTTING
FIXATION
TRIMMING (MICROTOMY)
DECALCIFICATION STAINING
(optional)
EMBEDDING
DEHYDRATION MOUNTING
INFILTRATION
(IMPREGNATION)
CLEARING LABELING


FIXATION
• prevent autolysis and bacterial attack.
• fix the tissues so they will not change
their volume and shape during
processing.
• prepare tissue and leave it in a
condition which allow clear staining of
sections.
• leave tissue as close as their living
state as possible, and no small
molecules should be lost.

DEHYDRATION
• to remove fixative and water from
the tissue and replace them with
dehydrating fluid.
• dehydration from aqueous fixatives
is usually initiated in 60%-70%
ethanol, progressing through 90%-
95% ethanol, then two or three
changes of absolute ethanol before
proceeding to the clearing stage

CLEARING
• process of replacing the
dehydrating fluid with a fluid that is
totally miscible with both the
dehydrating fluid and the
embedding medium.
Some clearing agents:

Xylene, Toluene, Chloroform, Benzene

INFILTRATION
• The melted wax will penetrate in
between the cells of the tissues.
• This process of wax infiltration is a
necessary step to harden the
tissues before their embedding.
• Infiltrating/embedding medium
almost always a Paraffin

EMBEDDING
• is the process by which tissues
are surrounded by a medium
such as agar, gelatin, or wax
which when solidified will
provide sufficient external
support during sectioning.

TRIMMING
• The resulting paraffin block is trimmed to
expose the tissue for sectioning (slicing)
on a microtome.
SECTION CUTTING
• Process of sectioning biological specimens
into very thin segments for microscopic
examination by means of a mechanical
equipment, called the microtome.

STAINING
• makes the various tissue
components conspicuous and to
permit distinctions between them
• H & E stains are universally used
for routine histological
examination of tissue sections.
• Hematoxylin stains acidic
molecules shades of blue. Eosin
stains basic materials shades of
red, pink and orange.


References
Mescher, A. L. (2018). Junqueira’s basic histology: Text and atlas, fifteenth edition (15th ed.)., McGraw-Hill
Gregorios. J. H. B., & Faldas, M. M. (2016). Histopathologic techniques (2016 revised ed.). Katha Publishing Co.
Materials from CPU-CMLS’ previous Human Histology instructors.

MODULE 1
EPITHELIAL
TISSUE

Module 2 – Epithelial Tissue Student Learning Outcomes
2.1. Organization of the body system 1. Understand comprehensively the

2.1.1 Cells organization of cells, tissues, organs and
2.1.2 Tissues system
2.1.3 Organs 2. Understand completely the functions of the
2.1.4 System epithelial tissues
2.2. Functions of the epithelial tissues 3. Determine appropriately the embryonic
2.3. Embryonic Origin of the epithelial cell that origin of the epithelial tissues that covers
covers different parts of the body the different parts of the body
2.4. Morphological characteristics of the 4. Describe properly the morphological
epithelial cells characteristics of the epithelial cells
2.5. Classification of epithelial cells 5. Identify correctly the classifications of the
2.6. Surface modifications of the epithelial cells epithelial tissues
6. Determine properly the surface
modifications of epithelial cells

Organizations of the Body System
TISSUES
• Collections of specialized cells and the extracellular
substances surrounding them.
Classified based on the following:

• structure of the cells
• the composition of the noncellular substances
surrounding the cells
• functions of the cells

EPITHELIAL CONNECTIVE MUSCLE NERVOUS
Four Basic Types of Tissues


EPITHELIAL TISSUE

Epithelial Tissues and its functions
EPITHELIAL TISSUE
• Epi + thele
• upon & nipple
• line all external and internal surfaces of the
body
• Composed of closely aggregated polyhedral
cells and to a thin layer of ECM.
Functions:
• Covering, lining, and protecting surfaces
• Absorption
• Secretion

Embryonic Origin of Epithelial Cells

Embryonic Origin of Epithelial Cells
ECTODERM
• epithelium that covers the external surface of the body, skin, cornea of the eyes
ENDODERM
• epithelium that lines digestive tract, liver, gall bladder, pancreas, respiratory tract,
urinary bladder, urethra
• except mouth & anus (ectoderm)
MESODERM
• epithelium that lines the heart, blood, lymphatic vessels & serous cavities, urinary
system (kidneys except bladder), male & female reproductive system.

Characteristic Features of
Epithelial Cells
1. Shapes and dimensions are quite
variable
2. Indistinguishable by light microscopy
3. Avascular
4. Show polarity
5. Exhibit continuous renewal

Characteristic Features of Epithelial Cells
1. Shapes and dimensions are
quite variable
• tall columnar to cuboidal to low squamous
cells.
• The cells’ size and morphology are generally
dictated by their function.
• cell's nuclei vary in shape and may be elliptic
(oval), spherical, or flattened, with nuclear
shape corresponding roughly to cell shape.

2. Indistinguishable by light
microscopy
• lipid-rich membranes of epithelial cells
• the number and shape of their stained nuclei
are important indicators of cell shape and
density.
• the number of cell layers in an epithelium
• a primary morphologic criterion for
classifying epithelia.

3. Avascular
• cells receive nutrients and O2 by diffusion
from the connective tissue.
• Lamina propria
– connective tissue that underlies the
epithelia lining the organs of the
digestive, respiratory, and urinary
systems
• Papillae
‒ area of contact between the two tissues
‒ projecting from the connective tissue
into the epithelium.

4. Show polarity
• with organelles and membrane
proteins unevenly distributed in
the cell.
✓ basal pole
✓ apical pole
✓ lateral surface

5. Exhibit continuous renewal
• relatively labile structures
• mitotic activity and stem cell populations.
• The rate of renewal varies widely
• fast
‒ in tissues such as the intestinal
epithelium, which is replaced every week
• slow
‒ large glands (liver)

Basement Membrane
• a thin extracellular, felt-like sheet of
macromolecules where the basal surface of all
epithelia rests
• a semipermeable filter
• rich in glycoproteins & proteoglycans
• 2 layers:
✓ Basal lamina
✓ Reticular lamina

Basement Membrane
Basal Lamina
• external laminae
• Nearest the epithelial cells
• an electron-dense layer
• 20-100 nm thick
• consisting of a network of fine fibrils
• made up of the ff
• Type IV collagen
• Laminin
• Nidogen and perlecan

Basement Membrane
Reticular Lamina
• below the basal lamina
• more diffuse and fibrous
• contains type III collagen and type VII
collagen

Basement Membrane
Functions:
1. attach epithelia to connective tissue
2. regulate (filter) substances passing from
connective tissue into epithelia
3. provide a guide or scaffold during tissue
regeneration after injury
4. compartmentalize epithelial cells from
other tissues.

Intercellular Adhesion & Junctions
• mechanically bind the cells together
• help form a permeability barrier
• provide a mechanism for
intercellular communication.

Intercellular Adhesion & Junctions
1. Tight or occluding junctions
• form a seal between adjacent cells.
2. Adherent or anchoring junctions

• are sites of strong cell adhesion.
3. Gap junctions
• are channels for communication between
adjacent cells.

Specializations of the
Apical Cell Surface
• Microvilli
• Steroecillia
• Cilia

Specializations of the Apical Cell Surface
Microvilli
• small membrane projections with cores of
actin filaments
• specialized for absorption at the apical cell
surfaces
• a brush or striated border
• average microvillus is about 1-μm long and
0.1-μm wide
• the total surface area can be increased by
20- or 30-fold.

Stereocilia
• best seen on the absorptive epithelial cells
lining the male reproductive system
• increase the cells’ surface area, facilitating
absorption.
• specialized stereocilia with a motion-
detecting function are important
components of inner ear sensory cells.

Cilia
• long, highly motile apical structures
• containing internal arrays of microtubules
Primary cilium
• not motile but is enriched with receptors and signal
transduction complexes for detection of light, odors,
motion, and flow of liquid past the cells.
Motile cilia
• are abundant on cuboidal or columnar cells of many
epithelia.
• Typical cilia are 5-10 μm long and 0.2 μm
• made up of 9 + 2 assembly of microtubules
Types of Epithelia
• Covering (lining)
epithelia
• Secretory (glandular)
epithelia

Types of Epithelia
Covering (lining)
epithelia
• Cells of covering epithelia are
organized into one or more layers
that cover the surface or line the
cavities of an organ.

Classification of Covering Epithelia

Covering Epithelia
Simple Squamous Epithelium
STRUCTURE FUNCTION LOCATION
• Single layer • Diffusion • Lining of blood vessels
• flat often hexagonal cells • Filtration • the heart
• the nuclei appear as bumps • some secretion • lymphatic vessels
when viewed as a cross section • some protection against friction (endothelium)
because the cells are so flat • small ducts
• alveoli of the lungs
• portions of the kidney tubules
• lining of serous membranes
(mesothelium) of the body
cavities (pleural, pericardial,
peritoneal)
• inner surface of the tympanic
membranesMLS 2206 | HUMAN HISTOLOGY
Covering Epithelia
Stratified Squamous Epithelium
KERATINIZED NON-KERATINIZED
• packed with keratin filaments • with relatively sparse keratin
• found mainly in the epidermis of skin, where it • lines moist internal cavities
helps prevent dehydration from the tissue • mouth, esophagus, and vagina
• the flattened cells of the surface layer retain
their nuclei and most metabolic functions.

Covering Epithelia
Simple Cuboidal Epithelium
• Single layer of cube-shaped cells • Secretion and absorption • Kidney tubules glands and their
• some cells have microvilli (kidney • movement of particles ducts
tubules) or cilia (terminal embedded in mucus • choroid plexuses
bronchioles of the lungs) • lining of terminal bronchioles of
the lungs
• surfaces of the ovaries

Covering Epithelia
Stratified Cuboidal Epithelium
• Multiple layers of somewhat • Secretion and absorption • Sweat gland ducts,
cube-shaped cells • ovarian follicular cells,
• salivary gland ducts

Covering Epithelia
Simple Columnar Epithelium
• Single layer of tall, narrow cells • Movement of particles or for the • Glands and some ducts
• some cells have cilia (bronchioles movement of oocytes • bronchioles of the lungs
of lungs, auditory tubes, uterine • secretion by cells of the glands • auditory tubes
tubes, and uterus) or microvilli • absorption • uterus
(intestines) • uterine tubes
• stomach
• intestines
• gallbladder
• bile ducts
• ventricles of the brain

Covering Epithelia
Stratified Columnar Epithelium
• Multiple layers of tall, narrow • seen in the conjunctiva lining the • Conjunctiva
cells eyelids, where it is both protective
and mucus secreting.

Covering Epithelia
Pseudostratified Columnar Epithelium
• Single layer of cells • Synthesize and secrete mucus • Lining of the nasal cavity
• the nuclei of these cells are at onto the free surface • nasal sinuses
different levels and appear • move mucus (or fluid) • auditory tubes
stratified • pharynx
• the cells are almost always • trachea
ciliated • bronchi of the lungs
• associated with goblet cells that
secrete mucus onto the free
surface

Covering Epithelia
Transitional Epithelium
• Stratified cells that appear cube- • Accommodate fluctuations in the • Lining of the urinary bladder
shaped when the organ or tube volume of fluid in organs or • ureters
is not stretched and squamous tubes • superior urethra
when the organ or tube is • protect against the caustic • uterus
stretched by fluid effects of urine
• the number of layers also
decreases on stretch

Types of Epithelia
Secretory epithelia &
Glands
• Formed by cells specialized in fluid secretion.
• May synthesize, store and secrete proteins,
lipids or complexes of carbohydrate and
proteins

Secretory Epithelia & Glands
Secretory cells
• may synthesize, store, and release:
✓ proteins (eg. in the pancreas)
✓ lipids (eg. adrenal, sebaceous glands)
✓ complexes of carbohydrates and proteins
(eg. salivary glands).

Glands
• specialized organs
• commposed of secretory cells
• function mainly to produce and secrete
various macromolecules
• develop from an infolding or outfolding of
epithelium in the embryo

Formation of glands from covering epithelia. MLS 2206 | HUMAN HISTOLOGY
Glands
Exocrine
• glands remain connected with the surface
epithelium
• tubular ducts lined with epithelium which
deliver the secreted material where it is
used.
• contain contractile myoepithelial cells
• serve to help propel secretory products
from acini into the duct system

Glands
Endocrine
• glands that has no connection to their
original epithelium
• Ductless
• lack myoepithelial cells
• specialized either for protein or steroid
hormone synthesis
• hormones are transported in the blood to
target cells throughout the body.

Structural Classes of Exocrine Glands
SECRETORY
GLANDS
PORTIONS
Simple Tubular
• ducts not branched • either short or long
and coiled
Compound Acinar
• ducts with two or • rounded and
more branches saclike

Mechanism of Exocrine Gland Secretion
Merocrine secretion
• most common method of protein
or glycoprotein secretion
• involves typical exocytosis from
membrane-bound vesicles or
secretory granules.
• either serous or mucous
according to the nature of their
secretory products
• Classified according to the nature
of secretory products:
✓ Serous
✓ Mucous
Merocrine secretion
Serous Cells
• synthesize proteins that are mostly not
glycosylated, such as digestive enzymes.
• The cells have well-developed RER and Golgi
complexes and are filled apically with secretory
granules in different stages of maturation
• stain intensely with H&E
• Acini of the pancreas and parotid salivary glands

Merocrine secretion
Mucous cells
• Ex. goblet cells which contain mucins
• Mucins - heavily glycosylated protein
‒ form a layer of mucus when they
become hydrated
• typically larger than serous cells with flattened
basal nuclei
• also have RER and Golgi complexes and are filled
apically with secretory granules
• Stained with PAS
The simple columnar epithelium lining the
small intestine shows many isolated goblet
cells secreting mucus into the lumen.
Holocrine secretion
• cells accumulate product continuously as they
enlarge and undergo terminal differentiation
• culminating in complete cell disruption which
releases the product and cell debris into the
gland’s lumen.
• Ex. sebaceous glands

Apocrine secretion
• product accumulates at the cells’ apical ends
• portions of which are then extruded to release
the product together with small amounts of
cytoplasm and cell membrane.
• Ex. Mammary gland

References
Gonzales, E. G. (2014). Esteban and Gonzales’ Textbook of Histology (3rd ed..), C & E Publishing, Inc.

MODULE 1
INTRODUCTION


Module 1 - Introduction
1.1 – Introduction to Histology

1.2 – The Cell

Module 1.2 – The Cell Student Learning Outcomes
1.2.1. Meaning of a cell 1. Define properly the term cell

1.2.2. Function of a cell 2. State correctly the function of a cell
1.2.3. Parts of a cell 3. Illustrate and label precisely the parts of the
1.2.4. Function of each part of the cell cell
1.2.5. Structures of each part of the cell 4. Determine correctly the substances present
1.2.6. Substances present in the cell in the cell
1.2.7. Inclusions present in the cell 5. State precisely the function of each part of
1.2.8. Differences of endocytosis and exocytosis the cell
1.2.9. Microscopic technique in dealing with the 6. Demonstrate microscopically the structures
prepared slides (to be discussed in the lab) of each parts of the cell
7. Identify correctly the inclusion bodies
present in the cell
8. Identify correctly the cytoskeleton present
in the cytoplasm of the cell
9. Differentiate comprehensively endocytosis
from exocytosis.

Meaning of a Cell
The Cell
the basic units of all living things, including humans

Meaning of a Cell
DIFFERENTIATION
• The process by which cells of an
embryo become specialized
structurally to augment specific
cytoplasmic activities for functions
at the level of tissues and organs.

Functions of a Cell
• Cells metabolize and release energy.
• Cells synthesize molecules.
• Cells provide a means of communication.
• Cells reproduce and provide for inheritance.

Parts of a Cell

Function of each part of the cell | Structures of each part of the cell | Substances present in the cell | Inclusions present in the cell
CYTOPLASM
• homogenous substance that fills the cell and the various formed elements
embedded in it.

PLASMA MEMBRANE
• Outermost layer of the cell.
• 7.5-10 nm thick
• Lipid bilayer composed of phospholipids and cholesterol with proteins
extend across or are embedded in either surface of the lipid bilayer
• 45–50% lipids,
• 45–50% proteins
• 4–8% carbohydrates

Glycolipids
• include oligosaccharide chains that extend outward
from the cell surface and contribute to a delicate
cell surface coating to the glycocalyx
Integral proteins
• are incorporated directly within the lipid bilayer
• extracted only by using detergents to disrupt the
lipids.
Peripheral proteins
• bound to one of the two membrane surfaces,
particularly on the cytoplasmic side
• extracted from cell membranes with salt solutions

PLASMA MEMBRANE
Functions:
• outer boundary of cells
• controls the entry and exit of substances
• receptor proteins function in intercellular
communication
• marker molecules enable cells to
recognize one another
• catalyzes chemical reactions
• attachment for the cytoskeleton

Transmembrane Proteins &

Membrane Transport

Cell Transport Mechanism

Passive Membrane Transport Active Membrane Transport
The cell does not expend metabolic A mediated transport mechanism that
energy during transport. requires energy provided by ATP.
Diffusion, Facilitated diffusion, Osmosis Primary & Secondary Active Transport,
Vesicle Membrane Transport

Diffusion (Passive Membrane Transport)

• movement of a solute from an area of higher concentration to an area of lower
concentration within a solvent.
Channels
• are multi-pass proteins forming transmembrane pores through which ions or small
molecules pass selectively.
• Cells open and close specific channels for Na+, K+, Ca2+ and other ions in response
to various physiological stimuli. Water molecules usually cross the plasma
membrane through channel proteins called aquaporins.
Carriers
• are transmembrane proteins that bind small molecules and translocate them across
the membrane via conformational changes.

Facilitated
Diffusion
(Passive Membrane
Transport)

Osmosis (Passive Membrane Transport)

• movement of a solvent (water) across a selectively permeable membrane`from a
higher water concentration to lower water concentration
• Osmotic pressure
• Isosmotic solutions have the same concentration solute particles
• Hyperosmotic solutions have a greater concentration solute particles
• Hyposmotic solutions have a lesser concentration of solute particles

Osmosis (Passive Membrane Transport)

• In a hypotonic solution, cells swell and can
undergo lysis
• In an isotonic solution, cells neither swell nor
shrink
• In a hypertonic solution, cells shrink.

Primary Active Transport

• Active transport is a mediated transport
process that requires energy provided by
ATP.

Secondary Active Transport

• Involves the active transport of an ion, such as
sodium, and provides the energy necessary to
move a different ion or some other molecule into
the cell.
• For example, glucose moves from the lumen of the
intestine into epithelial cells by secondary active
transport .
• This process requires two transport proteins:
(1) A Na+–K+ pump actively moves Na+ out of the cell
(2) a carrier protein facilitates the movement of Na+
and glucose into the cell
Vesicular Transport
• Refers the movement of larger
volumes of substances across the
plasma membrane through the
formation or release of vesicles,
membrane-bound sacs, in the
cytoplasm

Vesicular Transport
Endocytosis is the movement of materials into cells
by the formation of a vesicle.
1. Phagocytosis
2. Pinocytosis
3. Receptor-mediated endocytosis

Vesicular Transport
Phagocytosis
• cell eating
• the movement of solid material
into cells
• macrophage & neutrophils

Vesicular Transport
Pinocytosis
• cell drinking
• the uptake of small droplets of liquids
and the materials in them.
transcytosis-accomplishes bulk transfer

of dissolved substances across the cell.

Vesicular Transport
Receptor-mediated
endocytosis
• involves plasma membrane receptors
attaching to molecules that are then
taken into the cell.

Vesicular Transport
Exocytosis is the secretion of
materials from cells by vesicle
formation.

Signal Reception & Transduction

Endocrine Signaling
• the signal molecules (here called hormones) are
carried in the blood from their sources to target cells
throughout the body.
Paracrine Signaling
• the chemical ligand diffuses in extracellular fluid but
is rapidly metabolized so that its effect is only local
on target cells near its source.
Synaptic signaling
a special kind of paracrine interaction,
neurotransmitters act on adjacent cells through
special contact areas called synapses
Autocrine signaling
• signals bind receptors on the same cells that
produced the messenger molecule.
Juxtacrine signaling
• important in early embryonic tissue interactions
• the signaling molecules are cell membrane–bound
proteins which bind surface receptors of the target
cell when the two cells make direct physical contact.
CYTOPLASMIC ORGANELLES
• metabolically active structures
• membranous or non-
membranous
• positioned by the cytoskeleton

CYTOPLASMIC ORGANELLES
• RIBOSOMES
• ENDOPLASMIC RETICULUM
• ROUGH, SMOOTH ER
• GOLGI APPARATUS
• SECRETORY GRANULES
• LYSOSOMES
• PROTEASOMES
• MITOCHONDRIA
• PEROXISOMES

Ribosomes
• are macromolecular machines
• 20 × 30 nm
• each a complex of rRNA and many
• proteins, attach to mRNA
• free or bound
• intensely basophilic
• stain intensely with hematoxylin and such
as methylene and toluidine blue
• Serves as site of protein synthesis

Endoplasmic Reticulum
• convoluted membranous network
• Series of membranes distributed
throughout the cytoplasm of the cell that
are continuation of the outer membrane of
the nuclear envelope
Cisternae
• encloses a series of inter-communicating
channel

Endoplasmic Reticulum
Functions:
• Synthesis: Provides a place for chemical reactions
• a. Smooth ER- lipid synthesis and
carbohydrate metabolism
• b. Rough ER -synthesizes proteins
• Transport: Moves molecules through cisternal
space from one part of the cell to another,
sequestered away from the cytoplasm
• Storage: Stores newly synthesized molecules
• Detoxification: Smooth ER detoxifies both drugs
and alcohol

Rough Endoplasmic Reticulum

• Membranes with parallel and flattened cisternae
• attached with ribosomes
• prominent in cells specialized for protein secretion
• pancreatic acinar cells (making digestive enzymes)
• fibroblasts (collagen)
• plasma cells (immunoglobulins).
• basophilic staining

Smooth Endoplasmic Reticulum

• Membranes with tubular & sac-like with
interconnected channels of various shapes and
sizes cisternae
• with no attached ribosomes
• less abundant
• SER is not basophilic and is best seen with the
TEM.
• Manufactures lipids and carbohydrates
• Detoxifies harmful chemicals & stores calcium

Golgi Apparatus
• many smooth membranous saccules, some
vesicular, others flattened
• all contain enzymes and proteins being
processed
• located near the nucleus.
• Modifies, packages, and distributes proteins and
lipids for secretion or internal use.
• present in larger numbers and is most highly
developed in cells that secrete protein (Ex.
salivary glands or the pancreas)

Secretory Granules
• Originated as condensing vesicles in the Golgi
apparatus,
• found in cells that store a product until its
release by exocytosis
• signaled by a metabolic, hormonal, or neural
message (regulated secretion).
• surrounded by membrane and contain a
concentrated form of the secretory product
• 200 times more concentrated than those in the
cisternae of the RER.
• intensely eosinophilic structures TEM of one area of a pancreatic acinar cell shows numerous
• Ex. zymogen granules mature, electron-dense secretory granules (S) in association
with condensing vacuoles (C) of the Golgi apparatus (G).
Lysosomes
• lysis-solution; soma- body
• membrane-limited vesicles
• spherical, diameter from 0.05 to 0.5 μm
• appear as uniformly granular, electron-dense
appearance in the TEM
• visualized by light microscopy after staining with
toluidine blue.
• sites of intracellular digestion and turnover of
In the TEM lysosomes (L) have a characteristic very electron-
cellular components. dense appearance and are shown here near groups of Golgi
cisternae (G). The less electron-dense lysosomes represent
heterolysosomes in which digestion of the contents is under
way. The cell is a macrophage with numerous fine cytoplasmic
extensions (arrows). (X15,000)

Lysosomes
• contain about 40 different hydrolytic enzymes
present in cells with phagocytic activity
• optimal activity at an acidic pH (~5.0) and
inactive at the pH of cytosol (~7.2)
• acid hydrolyases such as proteases, nucleases,
phosphatase, phospholipases, sulfatases, and β-
glucuronidase.
Cells in a kidney tubule show numerous purple lysosomes (L) in the

cytoplasmic area between the basally located nuclei (N) and apical ends
of the cells at the center of the tubule. Using endocytosis, these cells
actively take up small proteins in the lumen of the tubule, degrade the
proteins in lysosomes, and then release the resulting amino acids for
reuse. (X300) MLS 2206 | HUMAN HISTOLOGY
Lysosomes
Autophagy
• the removal of excess or nonfunctional
organelles
• enhanced in secretory cells that have
accumulated excess secretory granules and in
times of nutrient stress, such as starvation.
In this TEM the two autophagosomes at the upper left contain

portions of RER more electron dense than the neighboring normal
RER and one near the center contains what may be mitochondrial
membranes plus RER. Also shown is a vesicle with features of a
residual body (RB).
Proteasomes
• cylindrical structure made of four stacked rings
• composed of seven proteins including proteases.
• very small abundant protein complexes not associated
with membrane
• They function to degrade denatured or otherwise
nonfunctional polypeptides.
• deal primarily with free proteins as individual molecules
• located in the cytosol and nucleus

Mitochondria
• membrane-enclosed organelles with arrays of enzymes
specialized for aerobic respiration
• produces of adenosine triphosphate (ATP)
• spherical, rod-shaped, or threadlike structures; enclosed
by double membrane
• with diameters of 0.5-1 μm and lengths up to 10 times
greater.
• Abundant in cardiac muscle, cells of some kidney tubules

Mitochondria
• In light microscope, it appear as numerous
eosinophilic structures in H&E.
• Under the TEM, double membrane that together

create two compartments:
• innermost matrix
• narrow intermembrane space

Mitochondria
Outer membrane
• is sieve-like, containing many
transmembrane proteins called porins
that form channels through which small
molecules such as pyruvate and other
metabolites readily pass from the
cytoplasm to the intermembrane space.

Mitochondria
Inner membrane
• Cristae - many long folds that project into
the matrix and greatly increase the
membrane’s surface area.
• The number of cristae in mitochondria
also corresponds to the energy needs of
the cell.

Peroxisomes
• spherical organelles enclosed by a single membrane
• contains enzyme that produce & degrade hydrogen peroxide (H2O2)
• Oxidase - produces H2O2
• Catalase - immediately break down H2O2
• inactivate various potentially toxic molecules, particularly in the large and abundant
peroxisomes of liver and kidney cells.
• form in two ways:
• budding of precursor vesicles from the ER growth
• division of preexisting peroxisomes
• site for formation of bile acids and cholesterol

Peroxisomes
(a) By TEM peroxisomes (P) generally show
a matrix of moderate electron density.
Aggregated electron-dense particles
represent glycogen (G). (X30,000)
(b) Peroxisomes (P) in most species are
characterized by a central, more
electron-dense crystalloid aggregate of
constituent enzymes, as shown here.
(X60,000)
(c) A cultured endothelial cell processed by
immunocytochemistry shows many
peroxisomes (green) distributed
throughout the cytoplasm among the
vitally stained elongate mitochondria
(red) around the DAPI-stained nucleus
(blue).

THE CYTOSKELETON
• a complex array of
(1) Microtubules
(2) microfilaments (also called
actin filaments)
(3) intermediate filaments.
• determine the shapes of cells
• important role in the movements of
organelles and cytoplasmic vesicles,
and also allow the movement of
entire cells.

THE CYTOSKELETON
General Function of Cytoskeleton
1. Structural
• Provides structural support to cell
• stabilizes junctions between cells
2. Movement
• Assists with cytosol streaming and
cell motility
• helps move organelles and
materials throughout cell
• helps move chromosomes during
cell division MLS 2206 | HUMAN HISTOLOGY
THE CYTOSKELETON
General Function of Cytoskeleton
1. Structural
• Provides structural support to cell
• stabilizes junctions between cells
2. Movement
• Assists with cytosol streaming and
cell motility
• helps move organelles and
materials throughout cell
• helps move chromosomes during
cell division
Microtubules
• fine tubular structures within the cytoplasm
of all eukaryotic cells
• organized into larger, more stable arrays
called axonemes in the cytoplasmic
extensions called cilia and flagella.
• hollow, with an outer diameter of 25 nm
and a wall 5-nm thick
• confers significant rigidity to help maintain
cell shape
• vary in length, but can become many
micrometers long.

Microtubules
• form part of the system for intracellular
transport of membranous vesicles,
macromolecular complexes, and organelles.
• motor proteins – control the transport along
microtubules which use ATP in moving the
larger structures.
• Kinesins - anterograde transport
• cytoplasmic dyneins – retrograde
transport
• Important roles for this system include
extending the ER from the nuclear envelope to
the plasmalemma and moving vesicles to and
through the Golgi apparatus. MLS 2206 | HUMAN HISTOLOGY
Cilia
• Short, numerous membrane extensions supported by
large & stable microtubules (axonemes)
• project from the surface of cells and vary in number
from none to thousands per cell.
• Move materials over the surface of cells
Flagellum
• Extension of the plasma membrane containing of parallel
microtubules;
• Thicker than cilia (55 μm in length)
• In humans, propels spermatozoa

Centrosome
• the microtubule-organizing center for
the mitotic spindle and consists of
paired centrioles.
• contains two centrioles oriented
perpendicular to each other
• each about 0.2 μm in diameter and 0.3-
0.5 μm in length.
• composed of nine highly organized
microtubule triplets
• Determine cell polarity during cell
division

Function of each part of the cell | Structures of each part of the cell |
Substances present in the cell | Inclusions present in the cell
Microfilaments (Actin Filaments)

• composed of actin subunits and allow motility and
most contractile activity in cells, using reversible
assembly of the actin filaments and interactions
between these filaments and associated myosin family
proteins.
• thin (5-7 nm diameter), polarized polymers, shorter

and more flexible than microtubules
• Actin is very abundant in all cells, concentrated in

networks of actin filaments and in the cytoplasmic
region often called the cell cortex.
Microfilaments (Actin Filaments)

Functions:
• cytoplasmic streaming
• Transport of organelles, vesicles, and granules
• produce two cells by cytokinesis during mitosis
• produce the cell surface changes during endocytosis
• permit very forceful contractions in specialized cells
such as those of muscle

Microvilli
• Extension of the plasma
membrane containing
microfilaments
• Increase surface area of the
plasma membrane for
absorption and secretion
• modified to form sensory
receptors

Intermediate Filaments
• intermediate in size
• with a diameter averaging 10 nm
• confer increased mechanical stability to cell
structure
• made up of different protein subunits in
different cell types.
• coiled, rod-like dimers that form antiparallel
tetramers
• protofibrils-large cable-like bundles
A large and important class of intermediate filaments is composed
of keratin subunits, which are prominent in epithelial cells. Bundles
of keratin filaments called tonofibrils associate with certain classes
of intercellular junctions (J) common in epithelial cells and are
easily seen with the TEM, as shown here in two extensions in an
epidermal cell bound to a neighboringMLS cell.2206
(60,000X)
| HUMAN HISTOLOGY
Keratins (cytokeratins)
• Intermediate filaments of keratins form large
bundles (tonofibrils) that attach to certain
junctions between epithelial cells
• accumulate during differentiation in the
process of keratinization, producing an
outer layer of nonliving cells that reduces
dehydration.
• provides protection from minor abrasions
and produces various hard protective
structures of skin A large and important class of intermediate filaments is composed
of keratin subunits, which are prominent in epithelial cells. Bundles
of keratin filaments called tonofibrils associate with certain classes
of intercellular junctions (J) common in epithelial cells and are
easily seen with the TEM, as shown here in two extensions in an
epidermal cell bound to a neighboringMLS cell.2206
(60,000X)
| HUMAN HISTOLOGY
Vimentin
• the most common class III intermediate
filament protein
• found in most cells derived from embryonic
mesenchyme
• Desmin - found in almost all muscle cells
• glial fibrillar acidic protein (GFAP) -
found especially in astrocytes
A single cultured uterine cell stained fluorescently to reveal a

meshwork of intermediate filaments (green) throughout the
cytoplasm. Primary antibodies against the filament protein
desmin and fluorescein isothiocyanate (FITC)-labeled secondary
antibodies were used in the indirect staining technique, with the
nucleus counterstained blue with DAPI. (X650)

Neurofilament proteins
• Proteins of 3 distinct sizes
• Make heterodimers that form
the subunits of the major
intermediate filaments of
neurons.

Lamins
• a family of seven isoforms present in the cell
nucleus
• they form a structural framework called the
nuclear lamina just inside the nuclear
envelope
Bound to the inner membrane of the nuclear envelope is the

nuclear lamina, a meshwork assembled from lamins (class V
intermediate filament proteins).
INCLUSIONS
• cytoplasmic structures or
deposits filled with stored
macromolecules and are not
present in all cells.
• Lipid droplets
• Glycogen granules
• Pigmented deposits
• melanin
(a) Lipid droplets
• lipofuscin (b) Glycogen granules
• hemosiderin (c) Pigment deposits (PD)

NUCLEUS
• a large organelle, usually located near the center of the
cell.
• it consists of a nuclear envelope containing chromatin,
the mass of DNA and its associated proteins, with one
or more specialized regions of chromatin called
nucleoli.
• Houses the DNA that serves as the genetic material for
directing protein synthesis
• control center of the cell

NUCLEUS

An electron micrograph
obtained by freeze-
fractured cell shows the
two layers of the
nuclear envelope and
nuclear pores. The
fracture plane occurs
partly between the two
nuclear envelope
membranes (left) but
mostly just inside the
envelope with the
chromatin removed.
The size and
distribution of the
nuclear pore complexes
are clearly seen.
(X60,000)

Nuclear Envelope
• forms a selectively permeable barrier
between the nuclear and cytoplasmic
compartments.
• the envelope has two concentric membranes
(outer and inner) separated by a narrow (30-
50 nm) perinuclear space
• nuclear pore complexes bridges the inner
and outer nuclear membranes
• Nucleoporins - core proteins of a nuclear
pore complex; regulates the movement of
macro-molecules between nucleus and
cytoplasm.
Chromatin
• consists of DNA and all of the associated proteins
involved in the organization and function of DNA.
• 46 chromosomes (23 pairs)
• Chromatids - two identical chromatin units
• Histones – small basic proteins where DNA
associates with to be packaged within the nucleus
• Nucleosome - structural unit of DNA and
histones; produces the initial organization of free
double-stranded DNA into chromatin

Chromatin
Types:
Euchromatin
• is visible as finely dispersed granular material
• lightly stained basophilic
• neurons
Heterochromatin
• appears as coarse, electron-dense material
• intensely basophilic clumps in the light microscope.
• lymphocytes, Barr bodies An active nucleus typically has much diffuse, light-staining
euchromatin and smaller subdomains of electron-dense
heterochromatin (H), with many of these associated at the
periphery associated with the nuclear lamina. The more
heterogeneous electron-dense subdomain is the nucleolus
(N), the site of rRNA synthesis, and ribosomal subunit
assembly. (X25,000) MLS 2206 | HUMAN HISTOLOGY
Nucleolus
• spherical, highly basophilic subdomain of nuclei
• engaged in protein synthesis
• The intense basophilia to the presence of densely
concentrated ribosomal RNA (rRNA) that is
transcribed, processed, and assembled into ribosomal
subunits.
• active nucleolus reveals fibrillar and granular
subregions with different staining characteristics that
reflect stages of rRNA maturation

THE CELL CYLCE

• repeated cycles of macromolecular
synthesis (growth) and division
(mitosis)
• The regular sequence of events that
produce new cell
• During this time, the cell carries out the
metabolic activities necessary for life
and performs its specialized functions

THE CELL CYLCE

Four Distinct Phases:
G1
• the time gap between mitosis and the
beginning of DNA replication
• the longest and most variable part of
the cycle
• period of active RNA and protein
synthesis, including proteins controlling
progress through the cell cycle.
• return of the cell volume

THE CELL CYLCE

S
• DNA replication, histone synthesis, and
the beginning of centrosome
duplication.
G2
• the gap between DNA duplication and
the next mitosis
• proteins required for mitosis accumulate

THE CELL CYLCE

G0
• cell cycle activities may be temporarily
or permanently suspended
• Cycling is activated in postmitotic G0
cells by protein signals from the
extracellular environment called
mitogens or growth factors that bind to
cell surface receptors and trigger a
cascade of kinase signaling in the cells.

MITOSIS
• The period of cell division
• parent cell divides and each of the two
daughter cells receives a chromosomal set
identical to that of the parent cell.
• Interphase is the phase between cell divisions
(the G1, S, and G2 phases).

STEM CELLS & TISSUE RENEWAL

• Undifferentiated cells
• cycling serves to renew the differentiated cells of
tissues as needed
• Stem cells divide asymmetrically. One daughter
cell remains as a stem cell, while the other
becomes committed to a path that leads to
differentiation (progenitor cells or “transit
amplifying cells”)
• blood cells, skin cells, and cells lining the
digestive tract

MEIOSIS
• a specialized process involving
two unique and closely
associated cell divisions that
occurs only in the cells that will
form sperm and egg cells.

MITOSIS MEIOSIS
Mitosis is a cell division that produces Meiosis involves two cell divisions and
two diploid cells. produces four haploid cells.
During meiotic crossing over, new

Lacking synapsis and the opportunity for
combinations of genes are produced
DNA recombination, mitosis yields two
and every haploid cell is genetically
cells that are the same genetically.
unique.

APOPTOSIS
• process of cell suicide
• rapid, highly regulated cellular activity that shrinks
and eliminates defective and unneeded cells
• It results in small membrane-enclosed apoptotic
bodies, which quickly undergo phagocytosis by
neighboring cells or cells specialized for debris
removal.

APOPTOSIS
Bcl-2 family
• cytoplasmic proteins that control apoptosis
• regulate the release of death-promoting factors
from mitochondria.
• activated by either external signals or irreversible
internal damage
TEM of a cell in late apoptosis shows radical changes in cell

shape, with membrane blebbing and the formation of many
membrane-bound cytoplasmic regions. These apoptotic bodies
may separate from one another but remain enclosed by plasma
membrane so that no contents are released into the
extracellular space. The membrane changes are recognized by
neighboring cells, and macrophages and apoptotic bodies are
very rapidly phagocytosed. (X10,000)
APOPTOSIS
Morphologic Features of cells undergoing Apoptosis:
1. Loss of mitochondrial function and caspase activation.
2. Fragmentation of DNA.
3. Shrinkage of nuclear and cell volumes
4. Cell membrane changes
5. Formation and phagocytic removal

References
Gregorios. J. H. B., & Faldas, M. M. (2016). Histopathologic techniques (2016 revised ed.). Katha Publishing Co.

MODULE 3
CONNECTIVE
AND
ADIPOSE TISSUE

Module 3 – Connective and Adipose Tissue
3.1 – Connective Tissue

3.2 – Adipose Tissue

Module 3.1
Connective Tissue
3.1 Connective tissues LO1: Determine correctly the

3.1.1. Importance importance of connective tissues
3.1.2. Functions
3.1.3. Types LO2: Understand comprehensively
3.1.4. Components the functions of connective tissues
3.1.5 Types of Fibers
3.1.6. Categories of Cells Present LO3: State correctly the different
3.1.7.Classifications types of connective tissues

3.1.1 Connective Tissues - IMPORTANCE
• Provides a matrix that supports and physically connects other tissues and
cells together to form the organs of the body
• Gives metabolic support to cells as the medium for diffusion of nutrients

and waste products.

3.1.2 Connective Tissues - FUNCTION
STRUCTURAL METABOLIC
1. Binding and packing material of 1. Vehicle for transport of gases and
other tissues substances to and from the various
tissues of the body
2. Forms the structural framework of 2. Venue for the exchange of gases

many organs and substances from the basic tissues
and the blood
3. Comprise the skeletal system 3. Storage area for fat

3.1.3 Types
Connective tissue
Embryonic Specialized
Connective Connective Connective
Tissue Tissue Proper Tissue
Mesenchyme Loose (areolar) Reticular CT Adipose Cartilage Bone Blood
Mucous Dense regular CT Hyaline Compact
Dense irregular Fibrous Spongy/Cancellous

CT
Elastic

3.1.4 Connective Tissues - COMPONENTS
• Major constituent is the Extracellular matrix (ECM)
• Which consists of different combinations of protein fibers and ground

substance.

COMPONENTS

GROUND SUBSTANCE IMPORTANCE
• Highly hydrated; transparent; • Allows diffusion of small
viscous molecules
• complex mixture of 3 major • Acts as lubricant and barrier to

kinds of macromolecules, the penetration of invaders
glycosaminoglycans (GAGs),
proteoglycans, and • Profoundly influence various
multiadhesive glycoproteins. cellular activities
• Filling the space between cells

and fibers in connective tissue
GROUND SUBSTANCE
• TEM of the connective tissue
shows GS as areas containing fine
granular material among collagen
fibers, elastic fibers, and
fibroblast process.

GROUND SUBSTANCE
• Glycosaminoglycans – mucopolysaccharides; long polymers of repeating
disaccharide units (hexosamine or uronic acid)
Hyaluronan (Hyaluronate or Hyaluronic acid)

• Largest and most ubiquitous GAG
• Important in allowing molecular diffusion through connective tissues and in
lubricating various organs and joints (binds a considerable amount of water)
Dermatan sulfate, Chondroitin sulfates, Keratan sulfate, and Heparan sulfate

• Major GAGs found in proteoglycans
• High negative charge – sequester cations such as water
GROUND SUBSTANCE
• Proteoglycans – consist of a core protein which are covalently
attached to sulfated GAGs
• Proteoglycans become bound to Hyaluronan by

a link protein and their GAG side chains associate
further with collagen fibers and ECM components
Perlecan – key proteoglycan in basal laminae

Aggrecan – large; core protein heavily bound with chondroitin and keratan
sulfate

GROUND SUBSTANCE
• Multiadhesive glycoproteins – large macromolecule with branched
oligosaccharide chains; multiple binding sites for cell surface integrins and for
other matrix macromolecules
Laminin – trimeric glycoprotein; binding sites for integrins, type IV collagen, and
specific proteoglycans = provide adhesion for epithelial cells
Fibronectin – dimer synthesized by fibroblast; binding sites for collagen and

specific GAGs = forms insoluble fibrillar networks throughout the CT
Fibronectin substrates provides specific binding sites for integrins = cell adhesion
and cellular migration through ECM
GROUND SUBSTANCE
• Interstitial fluid – water in the ground substance of CT; ion composition
similar to plasma; contains plasma proteins of low molecular weight that pass
through thin walls of capillaries
Hydrostatic pressure (blood) – caused by

the pumping action of the heart (forces
water out of the capillary wall
Colloid Osmotic pressure – produced by
plasma proteins (albumin) which draws
water back to the capillaries

3.1.5 Connective Tissues – TYPES OF FIBERS
COLLAGEN RETICULAR ELASTIC

FIBER FIBER FIBER

COLLAGEN
• Key element of all CT, epithelial basement membranes, and external laminae
of muscle and nerve cells
• Most abundant protein in human body (30% of its dry weight)
FUNCTION
• Able to form various extracellular fibers, sheets, and networks, all of which
are extremely strong and resistant to normal tearing and shearing forces

COLLAGEN
Collagen fiber
400x,H&E
COLLAGEN - CLASSIFICATION
• FIBRILLAR COLLAGEN – have polypeptide subunits that aggregate to form
large fibrils
*Type I, II, III, V, and IX
• NETWORK OR SHEET-FORMING COLLAGENS – subunits produced by
epithelial cells and major structural proteins of the external laminae and all
epithelial basal laminae
*Type IV and X
• LINKING/ANCHORING COLLAGENS – short collagens that link fibrillar
collagens to one another and to other components of the ECM
*Type VII, IX, XII, and XIV

• FIBRILLAR COLLAGEN
TYPE DESCRIPTION LOCATION MAIN FUNCTION
I Thick, highly picrosirius Skin, tendon, bone, dentin Resistance to tension

birefringent, fibers
II Loose aggregates of fibrils, Cartilage, vitreous body Resistance to pressure
birefringent
III Thin, weakly birefringent, Skin, muscle, blood vessels Structural maintenance in
argyrophilic (usually together with type I) expansible organs
V Frequently forms fiber Fetal tissues, skin, bone, Participates in type I collagen
together with type I placenta, most interstitial tissues function
XI Small fibers Cartilage Participates in type II collagen
function
• NETWORK-FORMING COLLAGEN
IV Two-dimensional cross-linked All basal and external laminae Support of epithelial cells;
network detected by filtration
immunocytochemistry
X Hexagonal lattices detected Hypertrophic cartilage involved in Increases density of the matrix
by immunocytochemistry endochondral bone formation

• LINKING/ANCHORING COLLAGEN
VII Detected by Epithelial basement membranes Anchors basal laminae to

immunocytochemistry underlying reticular lamina
IX Detected by Cartilage, vitreous body Binds various proteoglycans;
immunocytochemistry associated with Type II Collagen
XII Detected by Placenta, skin, tendons Interacts with Type I collagen
immunocytochemistry
XIV Detected by Placenta, bone Binds with Type I collagen fibrils,
immunocytochemistry with Types V and XII,
strengthening fiber formation

COLLAGEN SYNTHESIS

COLLAGEN SYNTHESIS

COLLAGEN DEGRADATION

RETICULAR
• Found in delicate CT of many organs (immune system)
• Consists mainly of Collagen Type III (forms an extensive network “reticulum”
of thin fibers
• Seldom visible in H&E; stained by impregnation of silver salts
FUNCTION
• Serve as supportive stroma for the parenchymal secretory cells and rich
microvasculature of the liver and endocrine glands
• Support rapidly changing populations of proliferating cells and phagocytic
cells
RETICULAR
Reticular fiber – Adrenal Cortex
100x, Silver-stained
ELASTIC
• Thinner than type I collagen fibers; form sparse networks interspersed with
collagen bundles especially in organs subject to regular stretching or bending
• Not strongly acidophilic and stain poorly with H&E; stain more darkly than
collagen with other stains such as orcein and aldehyde fuchsin
FUNCTION
• Allows tissue to be stretched or distended and return to their original shape

ELASTIC
Elastic fiber
200x, Hematoxylin-orcein
3.1.6 Connective Tissues – CATEGORIES OF CELLS
Components

FIBROBLAST ADIPOCYTES MACROPHAGES
LEUKOCYTES
MAST CELLS PLASMA CELLS MLS 2206 | HUMAN HISTOLOGY
RESIDENT (FIXED)
• Cells that permanently stay in the CT
• Fibroblasts and fibrocytes, reticular cells, adipose cells, mast cells, and
resident macrophage
VISITING (WANDERING)
• Cells that are transient in the CT
• Inflammatory macrophages, plasma cells, leukocytes

FIBROBLAST
• Most common cell in CT; produce and maintain most of the tissues
extracellular components
Function
• Synthesize and secrete collagen and elastin = both form large fibers (GAGs,
proteoglycans, and multiadhesive glycoproteins)
• Target of growth factors – influence cell growth and differentiation
• Involved in wound healing (myofibroblasts) – well developed contractile
function

FIBROBLAST
Fibroblast vs Fibrocyte
Fibroblast – active; more abundant and irregularly branched cytoplasm, more

RER, well-developed Golgi apparatus, large ovoid euchromatic nucleus, and
prominent nucleolus
Fibrocyte – quiescent; smaller, spindle-shaped with fewer processes, less RER,

darker, more heterochromatic nucleus

FIBROBLAST
Fibroblast Fibrocyte
400x,H&E 400x, H&E

ADIPOCYTES
• Fat cells; large mesenchymally derived cells
• Have major metabolic significance and considerable metabolic significance
Function
• Specialized for cytoplasmic storage of lipid as “neutral fats”
• Serves to cushion
• Insulate the skin and other organs

ADIPOCYTES
Adipocytes
100x, H&E

MACROPHAGES
• Size and shape vary considerably (state of functional activity)
• 10-30µm in diameter; eccentrically located oval or kidney-shaped nucleus
• Present in the CT of most organs, “Histiocytes”
Function
• Highly developed phagocytic activity
• Specialize in turnover of protein fibers and removal of apoptotic cells/tissue
debris (especially abundant in sites of inflammation)
• Secrete growth factors important for tissue repair
• Important in the uptake, processing, and presentation of antigens for
lymphocyte activation
MACROPHAGES
Macrophage
10,000x, TEM
Cells of the mononuclear phagocyte system

MACROPHAGES
Monocyte Macrophage
1,500x, Wright stain
researchgate.net/figure/Light-microscopic-analysis-of-human-monocyte-derived-macrophages-infected-with-S_fig1_14097817
MAST CELLS
• Oval, irregularly shaped; 7-20µm in diameter
• Filled with basophilic secretory granules that obscure the nucleus
Function
• Localized release of bioactive substances important in local inflammatory
response, innate immunity, and tissue repair

MAST CELLS
Mast cells
400x, Pararosaniline-Toluidine Blue stain

MAST CELLS – granules
• Electron dense; variable size (0.3 to 2.0 µm diameter)
• Display metachromasia – can change the color of some basic dyes (Toluidine
blue) from blue to purple to red
• Due to the high content of acidic radicals in their sulfated GAGs
• Poorly preserved by common fixatives – difficult to identify in routinely

prepared slides

MAST CELLS - bioactive substances
• Heparin – sulfated GAG; anticoagulant
• Histamine – promotes increased vascular permeability and smooth muscle
contraction
• Serine proteases – activate various mediators of inflammation
• Eosinophil and neutrophil chemotactic factors – attract leukocytes
• Cytokines – polypeptides directing activities of leukocytes and other cells of
the immune system
• Phospholipid precursors– converted to prostaglandins, leukotrienes, and
other important lipid mediators of the inflammatory response

MAST CELLS

PLASMA CELLS
• B Lymphocyte-derived, antibody-producing cells
• Large ovoid cells with basophilic cytoplasm rich in RER and large Golgi
apparatus near the nucleus
• Nucleus – spherical; eccentrically placed; contain compact, peripheral regions
of heterochromatin alternating with lighter areas of euchromatin
• Lifespan: 10-20 days
Function
• Synthesis of immunoglobulin antibodies

PLASMA CELLS
Plasma cells in infected lamina propria

Plasma cells
400x, Pararosaniline Toluidine Blue
640x, H&E
LEUKOCYTES
• White blood cells; derived from circulating wbcs
• Most wbcs stay in the CT for a few hours or days then undergo apoptosis
Function
• Vascular and defensive response to injury or foreign substances including
pathogenic bacteria or irritating chemical substances

LEUKOCYTES

3.1.7 Connective Tissues – CLASSIFICATION

EMBRYONIC CT
• “All connective tissues originate from the embryonic mesenchyme”
MESENCHYME
• Tissue developing mainly from the middle layer of the embryo (mesoderm)
• Consists largely of viscous substance with few collagen fibers
Mesenchymal cells – undifferentiated and have large nuclei; prominent nucleoli

and fine chromatin
“spindle-shaped” – scant cytoplasm extended as two or more thin cytoplasmic
processes

EMBRYONIC CT
MUCOID (MUCOUS) CT
• Principal component of the fetal umbilical cord (Wharton’s jelly)
• Abundant Ground substance composed of hyaluronan
• Gelatinous with sparse collagen fibers and scattered fibroblasts
• Similar to the tissue found in the vitreous chambers of eyes and pulp cavities
of young teeth

EMBRYONIC CT
Section of the umbilical cord
Mesenchyme (Mesenchymal cells) 200x, H&E
200x, Mallory trichrome

CONNECTIVE TISSUE PROPER
LOOSE CONNECTIVE (AREOLAR)
• Typically contains cells, fibers, and ground substance in roughly equal parts
• Common
• Fibroblast – most numerous cells; Collagen fibers predominate

• Has delicate consistency; Flexible; not resistant to stress
• FUNCTION
• forming a layer beneath epithelial lining of many organs and filling the spaces
between fibers of muscle and nerve

LOOSE CONNECTIVE (AREOLAR)
Skin
100x, Mallory trichrome

DENSE REGULAR CT
• Consist mainly of Type I collagen bundles and fibroblasts aligned in parallel
• Consist densely packed collagen separated by very little ground substance
• EXAMPLES : tendons (cords connecting muscles to bones); aponeuroses

(sheetlike tendons); ligaments (bands or sheets that hold together
components of the skeletal system
• FUNCTION
• great resistance to prolonged and repeated stresses from the same direction
DENSE REGULAR CT
Tendon
100x, H&E

DENSE IRREGULAR CT
• Bundles of collagen fibers appear randomly interwoven with no definite
orientation
• EXAMPLES : deep dermis layer of skin and capsules surrounding most organs
• FUNCTION
• Protects organs and strengthens them structurally

DENSE IRREGULAR CT
Gland
100x, H&E

SPECIALIZED CT
RETICULAR CT
• Characterized by abundant fibers of type III collagen forming a delicate
network that supports various types of cells.
• Macrophages and dendritic cells are also dispersed within these reticular
tissues to monitor cells formed there or passing through and to remove
debris
• EXAMPLES : Peyer’s patches, Spleen, Kidney, Bone Marrow
• FUNCTION
• Provides a framework with specialized microenvironment for hematopoietic
tissues in BM and lymphoid organs MLS 2206 | HUMAN HISTOLOGY
SPECIALIZED CT
RETICULAR CT
Lymph node
200x, Silver

SPECIALIZED CT
ADIPOSE TISSUE
• fat-storing cells or adipocytes predominate
• Types: White adipose tissue and Brown adipose tissue
• FUNCTION
• Storage depots for neutral fats (triglycerides)
• Provides thermal insulation for the body
• help shape the body surface, and cushion regions subject to repeated
mechanical stress such as the palms, heels, and toe pads

SPECIALIZED CT
ADIPOSE TISSUE
WHITE ADIPOSE TISSUE
• Unilocular; Signet ring appearance because the growing lipid droplets flattens
the nucleus of the adipocytes to the side
• Spherical when isolated but are polyhedral when closely packed in situ
BROWN ADIPOSE TISSUE

• Contain many small lipid inclusions and are therefore called multilocular
• Adults – found only in scattered areas – around kidneys, adrenal glands,
aorta, and mediastinum
• polygonal and generally smaller than white adipocytes; their smaller lipid
droplets allow the nucleus to be more centrally located MLS 2206 | HUMAN HISTOLOGY
SPECIALIZED CT
ADIPOSE TISSUE

SPECIALIZED CT
CARTILAGE
• Tough; durable form of supporting connective tissue
• Has firm consistency; resilient, smooth, lubricated surface
• Consists of CHONDROCYTES (Gr. Chondros, cartilage + kytos, cell) in ECM
• Contains no other cell types
• FUNCTION
• Allows tissue to bear mechanical stress without permanent distortion
• Provides cushioning and sliding regions within skeletal joints
• Facilitates bone movement
• Guides development and growth of long bones before and after birth
SPECIALIZED CT
CARTILAGE
• Lack vascular supplies
• Chondrocytes receive nutrients by diffusion from capillaries; avascular tissue;
low metabolic activity; lacks nerves
• Perichondrium – sheath of dense CT surrounds cartilage in most places;

harbors blood supply

SPECIALIZED CT
CARTILAGE

SPECIALIZED CT
CARTILAGE - Hyaline
• Most common; homogenous; semitransparent
• Adults – articular surfaces of movable joints, walls of larger respiratory

passages (nose, larynx, trachea, bronchi), ventral ends of ribs, epiphyseal
plates of long bones
• Embryo – forms the temporary skeleton that is gradually replaced by bone

SPECIALIZED CT
CARTILAGE - Hyaline
200x, H&E

SPECIALIZED CT
CARTILAGE - Elastic
• Similar to hyaline cartilage – contains an abundant network of elastic fibers in
addition to a meshwork of collagen type II fibrils
• More flexible than hyaline cartilage
• Found in the auricle of the ear, walls of the external auditory canals, the
auditory (Eustachian) tubes, epiglottis, URT

SPECIALIZED CT
CARTILAGE - Elastic
160x, Hematoxylin and Orcein

SPECIALIZED CT
CARTILAGE - Fibrous
• Takes various forms in different structures – essentially mingling of hyaline
cartilage and dense CT
• Found in intervertebral discs, attachments in certain ligaments, pubic

symphysis
• Serves as very tough yet cushioning support for the bone

SPECIALIZED CT
CARTILAGE - Fibrous
250x, Picrosirius-hematoxylin
SPECIALIZED CT
BONE
• Main constituent of adult skeleton
• Confers metabolic and mechanical functions to the skeleton
• Metabolites are unable to diffuse through the calcified matrix of bone, the
exchanges between osteocytes and blood capillaries depend on
communication through the very thin, cylindrical spaces of the canaliculi.
• FUNCTION
• Provides solid support for the body
• Protects vital organs such as those in the cranial and thoracic cavities
• Encloses internal (medullary) cavities containing bone marrow where blood
cells are formed MLS 2206 | HUMAN HISTOLOGY
SPECIALIZED CT
BONE
• 3 major cell type: Osteocytes, Osteoblasts, and Osteoclasts

SPECIALIZED CT
BONE - Compact
• Cortical bone; represents 80% of total bone mass
BONE - Cancellous
• Trabecular bone; 20% of total bone mass
• Deeper area with numerous interconnecting cavities

SPECIALIZED CT
BONE
Cancellous bone
250x
Compact bone
250x MLS 2206 | HUMAN HISTOLOGY

SPECIALIZED CT
BLOOD

Module 3 – Connective and Adipose Tissue
3.1 – Connective Tissue

3.2 – Adipose Tissue

Module 3.1
Connective Tissue
3.2 Adipose Tissue LO1: Determine correctly the

3.2.1. White Adipose Tissue importance of connective tissues
3.2.1.1. Storage and
Mobilization of Lipids LO2: Understand comprehensively
3.2.1.2. Histogenesis of White the functions of connective tissues
Adipose Tissue
3.2.2. Brown Adipose Tissue LO3: State correctly the different
3.2.2.1. Function of Brown types of connective tissues
Aipocytes
3.2.2.2. Histogenesis of Brown
Adipose Tissue

3.2 Adipose Tissues - INTRODUCTION
• CT where fat-storing cells/adipocytes predominate
• Cells are usually found isolated or in small groups within loose or dense
irregular CT
• Occur in large aggregates in adipose tissue or fat in many organs and body
regions
• Represents 15-20% of body weight in men, more in women
FUNCTION
• Storage depots for neutral fats
• Key regulator of body’s overall energy metabolism

3.2 ADIPOSE TISSUE
STORAGE
• Triglycerides
• Properties : insoluble in water – can be concentrated with no adverse
osmotic effects on cells
• Caloric density of TAGs – twice than that of carbohydrates and proteins as
well as glycogen
• Simple lipids are considered the most efficient means of storing calories

3.2 ADIPOSE TISSUE
ADIPOCYTES
• Active cells metabolically
• Respond to both nervous and hormonal stimuli
• Release hormones and other substances
Adipose tissue is now recognized as an endocrine gland at the center of
nutritional homeostasis.

3.2 ADIPOSE TISSUE
2 TYPES:
• WHITE ADIPOSE TISSUE
• BROWN ADIPOSE TISSUE

3.2.1 WHITE ADIPOSE TISSUE
• Most common type; specialize in fat storage

• Consists of cells each containing one large cytoplasmic droplet of whitish-
yellow fat
• Adipocytes - spherical when isolated but are polyhedral when closely packed
in situ; unilocular
• Very large; 50-150 µm in diameter
• “Signet-ring” appearance
• Contains a single huge droplet of lipid filling almost the entire cell

3.2.1 WHITE ADIPOSE TISSUE
• White adipose tissue associated with different
organs appears histologically similar,
differences in gene expression has been
observed.
• Important in the medical risks of obesity
• Increased visceral adipose tissue raises the risk

of diabetes and cardiovascular disease,
whereas increased subcutaneous fat does not
• During starvation adipocytes can lose nearly all

their fat and become polyhedral or spindle-
shaped cells with only very small lipid droplets.
3.2.1.1 STORAGE AND MOBILIZATION OF LIPIDS
WHITE ADIPOCYTES – can store triglycerides derived from:
• Dietary fats brought to the cells via the circulation as Chylomicrons
• Lipids synthesized in the liver and transported in the blood with VLDL
• Free fatty acids ang glycerol synthesized by the adipocytes
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3.2.1.2 HISTOGENESIS OF WHITE ADIPOSE TISSUE
• Adipocytes develop from mesenchymal stem cells.
• Adipose development first produces preadipocytes (look like larger fibroblast

with cytoplasmic lipid droplets)
• White adipocytes develop together with a smaller

population of cells termed beige adipocytes, which remain
within white adipose tissue and have histological and
metabolic features generally intermediate between white
and brown adipocytes.

3.2.1.2 HISTOGENESIS OF WHITE ADIPOSE TISSUE
• Humans are born with stores of white adipose tissue = begin to accumulate
by the 14th week of gestation.
• Proliferation of progenitor cells diminishes by late gestation
• Adipose tissue increases mainly by the filling of existing adipocytes until
around age 10
• New adipocyte formation occurs around small blood vessels, where
undifferentiated mesenchymal cells are most abundant.
• Excessive adipose tissue accumulation (obesity) - nutritional intake exceeds

energy expenditure (sedentary lifestyle)
• Weight loss after dietary changes is due to reductions in adipocyte volume,

but not their overall number.
3.2.2 BROWN ADIPOSE TISSUE
• Cells with multiple lipid droplets interspersed among abundant mitochondria;
darker appearance; multilocular
• Adipocytes – polygonal; generally smaller than white adipocytes

• Smaller lipid droplets allow the nucleus to be more centrally located
• Closely packed around large capillaries
• Receive direct sympathetic innervation to regulate their metabolic activity

3.2.2.1 FUNCTION OF BROWN ADIPOSE TISSUE
• Produce heat by nonshivering thermogenesis
• Heat production in brown adipocytes is greater than that of other cells
because their inner mitochondrial membranes have greatly upregulated
levels of the transmembrane protein uncoupling protein-1 (UCP1) or
thermogenin.
• Constitutes 2-5% of the newborn body weight located mainly at the back,
neck, and shoulders, but greatly reduced during childhood and adolescence
• Adults – found scattered around kidneys, adrenal glands, aorta, and

mediastinum

3.2.2.2 HISTOGENESIS OF BROWN ADIPOSE TISSUE
• Also develops from mesenchyme, but involves preadipocytes in a different
embryonic location
• Emerge earlier than white fat during fetal
development
• The number of brown adipocytes increases during
cold adaptation, usually appearing as clusters of
multilocular cells in white adipose tissue.
• As indicated earlier this increase involves the

reversible shift of beige cells to functional brown
adipocytes, but may also include proliferation and
differentiation of new adipocytes from preexisting
progenitor cells.

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MODULE 1

MLS 2206 | Human Histology

MLS 2206 | HUMAN HISTOLOGY

1.1 – Introduction to Histology

MLS 2206 | HUMAN HISTOLOGY

1.1.1. Meaning of Histology 1. Understand comprehensively the field

• the study of the tissues of the body and

• Involves all aspect of tissue biology, with

EPITHELIAL CONNECTIVE MUSCLE NERVOUS

Four Basic Types of Tissues

Cloudy swelling of the renal tubules. The outline of the

MLS 2206 | HUMAN HISTOLOGY

• Involve different procedures that have been

AIM OF HISTOPATHOLOGICAL TECHNIQUE:

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

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Some clearing agents:

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MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

Materials from CPU-CMLS’ previous Human Histology instructors.

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

2.1. Organization of the body system 1. Understand comprehensively the

MLS 2206 | HUMAN HISTOLOGY

Classified based on the following:

MLS 2206 | HUMAN HISTOLOGY

EPITHELIAL CONNECTIVE MUSCLE NERVOUS

Four Basic Types of Tissues

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

MLS 2206 | HUMAN HISTOLOGY

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2. Adherent or anchoring junctions

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