INTRODUCTION TO

HISTOLOGY
Prepared by
Dr. Belqees A. Allaw
2021
 Introduction
• Human anatomy consists of two main
divisions:
1- Macroscopic or gross anatomy
2- Microscopic anatomy (Histology)
vHistology is the science of the
microscopic structure
of cells, tissues and organs as seen
through a microscope.
• It also helps us understand the
relationship between structure and
function.
 Types of microscopes
1- The Compound Light Microscope
• one of the most familiar microscope as it is the
most often found in science and biology classrooms;
simple models and inexpensive.
• Commonly binocular (two eyepieces), the eyepiece
itself allows for 10X or 15X magnification.
• the three or four objective lenses, produce higher
magnification to a maximum of around 1000X
• Uses:
1- studying plant cells,
2- view bacteria and parasites as well as a variety of
human/animal cells.
3- useful in forensic labs for identifying drug
structures.
 Types of microscopes
2- The Stereo microscope (dissecting microscope)
it has two optical paths at slightly different angles
allowing the image to be viewed three-dimensionally
under the lenses.
• They magnify at low power, typically between 10X
and 200X, generally below 100x.
• Uses:
1- looking at surfaces.
2- microsurgery.
2- watch making, plus building and inspecting circuit
boards.
3- allow students to observe plant photosynthesis in
action.
 Types of microscopes
3- The digital microscope:
it invented in Japan in 1986, uses the power of the
computer to view objects not visible to the naked
eye.
• this kind can be found with or without eyepieces
to peer into.
• The computer software allows the monitor to
display the magnified specimen.
• Its advantage:
1- the ability to email images & recorded or
captured in the computer’s memory.
2- comfortably watch moving images for long
periods.
 Types of microscopes
4- The Electron Microscope
it is a powerful microscope available that allowing
researchers to view a specimen at nanometer size.
Its types:
1- The transmission electron microscope(TEM):
is capable of producing images 1 nanometer in size.
2- The scanning electron microscope(SEM):
approximately 10 times less powerful than TEMs,
they produce high-resolution, sharp, black and white
3D images.
Types of microscopes
• Both types have practical applications in:
1- fields as biology, chemistry, gemology,
metallurgy and industry.
2- provide information on the topography,
morphology, composition and crystallographic
data of samples.
 Anatomy of Light Microscope
The three basic, structural components of a
compound microscope are:
• Head/Body houses the optical parts in the
upper part of the microscope
• Base of the microscope supports the
microscope and houses the illuminator
• Arm connects to the base and supports the
microscope head. It is also used to carry the
microscope.
 Anatomy of Light Microscope
There are two optical systems in a compound
microscope:
1- Eyepiece or Ocular is what you look through at the top
of the microscope. Typically, standard eyepieces have a
magnifying power of 10x.
Eyepiece Tube holds the eyepieces in place above the
objective lens. Two types available: Binocular &
monocular microscope.
2- Objective Lenses are the primary optical lenses on a
microscope. They range from 4x-100x and typically,
include: three, four or five on lens in most microscopes.
 Anatomy of Light Microscope
• Nosepiece houses the objectives. Standard objectives
include 4x, 10x, 40x and 100x although different power
objectives are available.
• Coarse and Fine Focus knobs are used to focus the
microscope.
• Stage is where the specimen to be viewed is placed.
• Stage Clips are used when there is no mechanical stage.
• Aperture is the hole in the stage through which the base
light reaches the stage.
• Illuminator is the light source for a microscope.
Anatomy of Light Microscope
• Condenser is used to collect and focus
the light from the illuminator on to the
specimen.
• Iris Diaphragm controls the amount of
light reaching the specimen.
Definition
• Histology is the science of the
microscopic structure
of cells, tissues and organs as seen
through a microscope.
• It also helps us understand the
relationship between structure and
function.
 Types of microscopes
1- The Compound Light Microscope
• one of the most familiar microscope as it is the
most often found in science and biology classrooms;
simple models and inexpensive.
• Commonly binocular (two eyepieces), the eyepiece
itself allows for 10X or 15X magnification.
• the three or four objective lenses, produce higher
magnification to a maximum of around 1000X
• Uses:
1- studying plant cells,
2- view bacteria and parasites as well as a variety of
human/animal cells.
3- useful in forensic labs for identifying drug
structures.
 Types of microscopes
2- The Stereo microscope (dissecting microscope)
it has two optical paths at slightly different angles
allowing the image to be viewed three-dimensionally
under the lenses.
• They magnify at low power, typically between 10X
and 200X, generally below 100x.
• Uses:
1- looking at surfaces.
2- microsurgery.
2- watch making, plus building and inspecting circuit
boards.
3- allow students to observe plant photosynthesis in
action.
 Types of microscopes
3- The digital microscope:
it invented in Japan in 1986, uses the power of the
computer to view objects not visible to the naked
eye.
• this kind can be found with or without eyepieces
to peer into.
• The computer software allows the monitor to
display the magnified specimen.
• Its advantage:
1- the ability to email images & recorded or
captured in the computer’s memory.
2- comfortably watch moving images for long
periods.
 Types of microscopes
4- The Electron Microscope
it is a powerful microscope available that allowing
researchers to view a specimen at nanometer size.
Its types:
1- The transmission electron microscope(TEM):
is capable of producing images 1 nanometer in size.
2- The scanning electron microscope(SEM):
approximately 10 times less powerful than TEMs,
they produce high-resolution, sharp, black and white
3D images.
Types of microscopes
• Both types have practical applications in:
1- fields as biology, chemistry, gemology,
metallurgy and industry.
2- provide information on the topography,
morphology, composition and crystallographic
data of samples.
 Anatomy of Light Microscope
The three basic, structural components of a
compound microscope are:
• Head/Body houses the optical parts in the
upper part of the microscope
• Base of the microscope supports the
microscope and houses the illuminator
• Arm connects to the base and supports the
microscope head. It is also used to carry the
microscope.
 Anatomy of Light Microscope
There are two optical systems in a compound
microscope:
1- Eyepiece or Ocular is what you look through at the top
of the microscope. Typically, standard eyepieces have a
magnifying power of 10x.
Eyepiece Tube holds the eyepieces in place above the
objective lens. Two types available: Binocular &
monocular microscope.
2- Objective Lenses are the primary optical lenses on a
microscope. They range from 4x-100x and typically,
include: three, four or five on lens in most microscopes.
 Anatomy of Light Microscope
• Nosepiece houses the objectives. Standard objectives
include 4x, 10x, 40x and 100x although different power
objectives are available.
• Coarse and Fine Focus knobs are used to focus the
microscope.
• Stage is where the specimen to be viewed is placed.
• Stage Clips are used when there is no mechanical stage.
• Aperture is the hole in the stage through which the base
light reaches the stage.
• Illuminator is the light source for a microscope.
Anatomy of Light Microscope
• Condenser is used to collect and focus
the light from the illuminator on to the
specimen.
• Iris Diaphragm controls the amount of
light reaching the specimen.
Cell Morphology
 Cell biology

ØA cell is the smallest functional unit of an

organism.
• There are two general classes of cells:
1- Prokaryotes are the self-sustaining simple
cells (unicellular organisms) that characterized
by:
• lack organelles or other internal membrane-
bound structures.
• do not have a nucleus but instead generally
have a single chromosome.
• E.g: bacteria and archaea.
 Cell biology
2- Eukaryotic cells are more complex dependent
cells generally found in animals, plants, algae, and
fungi.
All cells of the human body are eukaryotic
(nucleus and cytoplasm).
ØThe cytoplasm contains specialized subunits
called organelles which work like ‘little organs.
• Organelles can be:
1- membranous (mitochondria, Golgi apparatus,
endoplasmic reticulum, nucleus)
2- non-membranous (ribosomes, nucleolus,
centrioles).
 Cell biology

vThe nucleus is a membrane-bound organelle

that contains genetic material (DNA:
deoxyribonucleic acid). it serves to maintain
the integrity of the cell by facilitating
transcription and replication processes.
vAll cells are enveloped by a semipermeable
two-layered membrane, which serves as a
dynamic medium for the cell’s interaction
with the external environment.
Components of Eukaryotic cell
ØPlasma (cell) membrane
It is the outermost layer of the cell which
consist of :phospholipid and protein
components.
• The main function of the plasma
membrane is to protect the cell from its
environment.
1- The cell membrane consists of a lipid
bilayer, including cholesterols that sit
between phospholipids to maintain their
fluidity at various temperatures.
Components of Eukaryotic cell
2- protein channels and carrier proteins anchored
in the plasma membrane that selectively or
generally allow particular particles to enter and
leave the cell.
• typically around 50% of membrane volume.
Membrane proteins consist of three main types:
1- integral proteins: ion channels
2- peripheral proteins: enzymes and hormones.
3- lipid-anchored proteins: G proteins (involved in
transmitting signals from a variety of stimuli
outside a cell to its interior).
Components of Eukaryotic cell
• Plasma membranes also contain
carbohydrates predominantly
glycoproteins.
• Carbohydrates are important in the
role of cell-cell recognition in
eukaryotes
 Components of Eukaryotic cell
• the plasma membrane has the ability to engulf
foreign material in a process known
as endocytosis which subdivided into three types:
1- Phagocytosis involves the intake of nonspecific
substances (usually solid) into the cell.
2- Pinocytosis involves the intake of specific
substances (usually extracellular fluid) into the
cell.
3- Receptor-mediated endocytosis involves the
specific uptake of certain macromolecules which is
controlled by cell surface receptors.
Components of Eukaryotic cell
ØCytoplasm:
• The semi-solid medium that keeps the organelles
suspended and nutrients dissolved within the
internal cellular environment.
• It contains microfilaments, microtubules and
secretory granules which:
1- helps give the cell its structure (cytoskeleton).
2- play a role in cell replication.
3- contribute to the formation of cilia and flagella in
some cell lines that require motility e.g: sperm.
Components of Eukaryotic cell
ØRibosomes
They are complex ribonucleic acid based molecules
(i.e. ribosomal-ribonucleic acid; r-RNA) that are
responsible for translating coded sequences of the
messenger-RNA (m-RNA) to proteins.
Types of Ribosomes:
1- Some ribosomes are membrane bound which
synthesize proteins that will be exported.
2- others float freely in the cytoplasm that synthesize
proteins that are used within the cell.
Components of Eukaryotic cell
ØEndoplasmic reticulum
clusters of sacs and vesicles that
form cisternae (tubules) within the cytoplasm.
There are two types of endoplasmic reticulum:
1- rough endoplasmic reticulum (RER).
2- smooth endoplasmic reticulum (SER).
 Rough ER Smooth ER
1- Shape ribosomes bound to its surface lacks ribosomes
2- Origin extension of the nuclear either be an independent
membrane collection of sacs, or a
continuation of the RER.
3- function stores protein that was synthesizes phospholipids,
synthesized by the ribosomes on steroids, and lipids which
its surface. subsequently used in steroid based
hormone synthesis.
Components of Eukaryotic cell
ØGolgi apparatus
• storage centre for proteins that will be
distributed to other sites.
• it is structurally subdivided
into cis and trans components.
1- cis components: are flattened incoming
vesicles from the endoplasmic reticulum that
fuse to form cisternae.
2- trans component: is the region from which
vesicles bud off to join other vesicles,
lysosome or the cell surface
Components of Eukaryotic cell
ØVesicles and lysosomes
They are membrane bound sacs to prevent some
proteins synthesised from being activated and
interacting unintentionally with the cell of origin.
ØThere are three general types of vesicles:
1- The exocytotic vesicles contain proteins that
will be expelled from the cell via exocytosis. e.g:
release of antibodies from activated B-cells during
the humoral immune response.
Components of Eukaryotic cell
2- secretory vesicles are also for
extracellular release but require a stimulus;
e.g: release of a neurotransmitter (ACh)
following stimulation by an action
potential.
3- lysosomes are stored enzymes that
designed to digest protein involved in an
apoptotic or as part of the defence
mechanism against invading pathogens.
Components of Eukaryotic cell
ØMitochondria
• the “powerhouse” of the cell.
• an elongated, double membrane structure
with numerous cristae within its inner
membrane.
• A crista is a fold in the inner membrane of a
mitochondrion
• They are the only organelles that contain
their own DNA material and is therefore
capable of replication.
Components of Eukaryotic cell
A- The outer membrane that envelops the entire
organelle that allow the selective uptake size of
some substances.
B- The inner membrane also has specific proteins
such as:
1- ATP synthase.
2- cytochrome C
3- transport proteins
The constituents of the intermembrane
space (between the inner and outer membranes)
are very similar to those in the cytoplasm of the
cell.
Components of Eukaryotic cell
C- The matrix is the site at which the
citric acid cycle (Krebs cycle - process in
ATP formation) occurs.
• The number of mitochondria found
within a particular cell is dependent
on its function.
E.g: cardiac myocytes contain more
mitochondria than epithelial cells of
the skin because they require more ATP
to make them resistant to fatigue.
 Components of Eukaryotic cell
ØNucleus
• the largest structure within the cell that
circumscribed by the nuclear envelope and
contains:
1- nucleolus.
2- matrix.
3- deoxyribonucleic acid (DNA): hereditary
genetic material.
DNA only exists as chromosomes during the
active stages of cellular division.
Components of Eukaryotic cell
üNucleolus is an area where DNA that codes for ribosomal RNA
that will be exported to the cytoplasm to translate m-RNA.
üNuclear envelope
• it is also made of a lipid bilayer.
• It is a double layered structure that encircles the nucleolus and
the chromatin within the nuclear matrix.
ünuclear pores: are openings in some areas of nuclear envelope
that:
1- allow nucleotides and other materials to enter the nucleus.
2- allow m-RNA to leave the nucleus for translation in the
cytoplasm.
 Plant cell VS Animal Cell
Plant cell Animal cell
fixed rectangular shape. irregular in shape
Cell wall : rigid cellulose-based cell wall that cell wall is absent.
surrounds the plasma membrane
Source of energy: ATP and chloroplasts. Source of energy is ATP only
missing cilia. They may be equipped with cilia.
How Histology Slides are
Prepared?
 1. Tissue Fixation
• its purpose is to prevent tissue autolysis and
putrefaction.
• most specimens are fixed in 10% neutral
buffered formalin.
• This will allow most tissues to become
adequately fixed within 24-48 hours.
• Formalin containers should be capped and
leak-proof, and labelled correctly.
2. Specimen transfer to cassettes
• specimens are trimmed using a scalpel to
enable them to fit into an appropriately
labelled tissue cassette.
• they must not be too thick (ideally should be
less than 4mm).
• stored in formalin until processing begins.
3. Tissue processing
• Processing tissues into thin microscopic sections is usually done
using a paraffin block as follow:
1- Dehydration: immersing your specimen in increasing
concentrations of alcohol to remove the water and formalin from the
tissue.
2- Clearing: an organic solvent such as xylene is used to remove the
alcohol and allow infiltration with paraffin wax.
3- Embedding: specimens are infiltrated with the embedding agent –
usually paraffin wax; creating “block”.
Once the block solidifies, it provides a support matrix that allows very
thin sectioning.
Tissue processing
 4. Sectioning
• Blocks are chilled on a refrigerated plate or ice
tray for 10 minutes before sectioning.
• A microtome is used to slice extremely thin tissue
sections off the block in the form of a ribbon;
most tissues are cut at around 5 µm.
• carefully transferred to a warm water bath that
help to be scooped up onto a slide placed under
the water level.
• dry upright at 37oC for a few hours to gently melt
the excess paraffin wax, leaving the tissue section
intact.
5. Staining
• Most cells are transparent, and appear almost
colourless when unstained.
• Histochemical stains (haematoxylin and eosin)
are used to provide contrast to tissue sections,
making tissue structures more visible and easier
to evaluate.
• Finally, a cover slip is mounted over the tissue
specimen on the slide, using optical grade glue,
to help protect the specimen.
Cells & tissue
vHematoxylin and eosin (H&E) stains are
one of the most common stains used in
histology and viewed under simple light
microscopy.
1- hematoxylin is a dark blue or violet stain
that is basic/positive and binds to basophilic
substances such as nucleic acid (round blue
structure)
2- Eosin stains eosinophilic structures in
shades of red, pink or orange, and will
highlight other structures of the cell.