Histology and Embryology

INTRODUCTION

I. What’s histology?

II. Why we study it ?

III. How to study it ?-Histological methods.

I. What’s histology?
Histology (Greek words):
/histo-tissue
/logia-study of ,or knowledge of histology means the
knowledge of tissue, is a branch of Anatomy.
Anatomy:
-gross anatomy
-microscopic anatomy
Structures related to function. So, exactly, Histology is a
science which study the microstructure and the
relationship between the structure and function of
human being.
Cytology –cytos –cell and logos science is science of the cell.
Histology –histo- tissue and logos science is science of the tissue.
Embriology- embryon and logos

It’s a complex of specialized cells, unated of structural, metabolic and

functional signs in more complex biological structure.

Histology is combination of two parts

1. General histology
2. Specialized histology or Tissue histology.
Cell: smallest unit of structure and function of body
↓
tissue: group of cell+extracellular ground substance
four basic tissue:
-epithelium
↓ -connective tissue
-muscular tissue
-nervous tissue
organ: made up of tissue, have special shape, structure
and function
↓
system: organs which have related function get together.
• It is the bases of other subject in medicine.

• It intertwines the disciplines of cell biology,

biochemistry, physiology, and as appropriate,
pathology. Students will recognize the
importance of this subject as they refer to the
text later in your careers.
II. What’s Embryology?

Embryology is a part of natural sciences which study the processes and

the regulations of the development of human fetus.
III. How to study it- histological methods
-Development of histology depends on the
development of technique.

-Histology studies the microstructures. So, we should

have the aid of microscope to study. Several types of
microscopes are available. According to the light
source used, microscopes can be basally classified as:
• light microscope(LM)
• electron microscope(EM)
1. Structure of Microscope
LM EM
-useful magnification: 1500X 800,000X
-resolution: 0.2um 0.2nm
2. Preparation of tissue for LM
The most routine one is paraffin section stained with
hematoxylin and eosin
The steps:
a. Obtaining the specimen: fresh, small pieces (less than
5 cubic millimeter (mm3))-tissue block
b. Fixation: fixatives: use formalin or Bouin’s to preserve
structural organization
Bouin's solution is a popular fixative for embryonic studies and skin, due
to its excellent preservation of nuclei and chromosomes. Bouin's is very
compatible with the trichrome stain due to its mordanting effect on the
tissue.
c. Dehydration: use ethyl alcohol to get rid of water of
tissue and cell
d. Clearing: use xylene to get rid of alcohol
alcohol and xylene are embedding mediums
e. Embedding: firstly, heat the paraffin, make it melt,
then put tissue block into melted paraffin, allow
paraffin harden, the tissue block is embedded in.
f. Sectioning: use microtome to cut the tissue into 3-
8 mm thick sections, then mounted them on glass
slides
g. Staining
-Hematoxylin: basic stain, combines with acidic
components, make them appear blue color- we call
such components as basophilic
-Eosin: acidic stain, combines with basic
components, make them appear pink color- we call
such components as acidophilic (eosinophilic)
observing
3. Preparation of tissue for EM
The steps are same to preparation for LM
a. tissue block: more small, less than 1mm3
b. plastic materials for embedding
c. ultra-thin sections is about 30-50nm thick( use
ultramicrotome)
d. heavy metal salts- increase staining contrast
-lead citrate
-uranyl acetate
 e. the beam of electron replace the light to
illuminate the tissue sections
Beam of electron illuminate the tissue section,
we use a screen to receive the electron. In some
areas, the beam of electron is impeded by those
tissue element which are stained with heavy metal
salts, so very few electrons penetrate to excite the
screen, such areas appear dark, are described as
electron-dense areas. Unstained areas, by contrast,
appear light, we call them as electron-lucent areas.
4. Histochemistry
1) General Histochemistry:
Combine histological methods with chemical or
biochemical methods, make some compositions of
tissue or cell become insoluble, colored or
electron-dense, to show those chemical
compositions of tissue or cell in situ, such
compositions includes protein, amino acid, nucleic
acid, lipid and enzymes.
*Periodic acid schiff reaction (PAS reaction):
schiff’s reagent (colorless)
Polysaccharides → aldehydes → magenta complexes
Glycogen oxidize combine (purple red colored)
3) in situ hybridization: nucleic acid: DNA (desoxyribose nucleic
acid) RNA (ribose nucleic acid)
Histology

It is the study of the tissues of the body and how

these tissues are arranged to constitute organs
also called Microscopic anatomy or
Microanatomy.
Preparation of tissue For Light Microscopy (
Tissue Processing)

• The aim of tissue processing is to embed the tissue in

a solid medium firm enough to support the tissue and
give it sufficient rigidity to enable thin sections to be
cut, and yet soft enough not to damage the knife or
tissue with preservation of the structure with the least
possible alteration.
Tissue Processing

Fixation
Dehydration
Clearing
Embedding
Sectioning
Sliding
Staining
Covering
Collecting the Sample
 Clinical Details
 Adequate specimen
Fixation

• To preserve the structure of tissue

• Achieved by the influence of various
chemical compounds called Fixative.
• Common methods - 10% formaldehyde
• Time require is almost 1mm/hour fixation
• It is also used protect the tissue from the
microorganisms
Dehydration
• Is the removal of extractable water from the tissue.
• Graduated strength of ethyl alcohol is routinely employed at
series consisting of 30,50,70,95 and 100% alcohol produce good
result.
• Average time required is 20-30 min in each solution
Clearing

• Also called dealcoholation.

• The aim of this process is to replace alcohol by a solvent which
is miscible with paraffin.
• Xylene and chloroform are the most commonly used.
Embedding

• Before sectioning of tissue it must be embedded in a material

which after hardening has a consistency that permits it to be cut
into thin section.
• Paraffin wax is the most frequency used agent.
• During embedding process paraffin heated to 60 degree, and in
cooling the paraffin with the tissue forms a firm tissue block.
Sectioning
• Sectioning is carried out by the help of machine called
Microtome.
• Usually 3-10 µm thick
Sliding
• Also called mounting of the slide.
• Clean microscopic glass slid are taken and the section which is
floated in warm water is taken on the glass slid in such a way
that no air bubble is trapped between them.
• The dish is always has black ground with rounded edge
Staining
• Because the tissue of the body are colourless and it’s difficult to
study their details the staining techniques enhance natural
contrast and permits distinction to be made between them.

• When the colouring property of the dye is in the basic radical

the stain is called basic dye and the stained structures called
Basophilic . On the other hand, When the colouring property of
the dye is in the acidic radical the stain is called acidic dye and
the stained structures called Acidophilic.
• The most commonly used dye is a combination of
HEMATOXYLIN and EOSIN
• To do it you must go in reverse order process
because this stain dissolve in water.
• By this method the nuclear structures are stained
dark purple or blue and all cytoplasmic structures
and intracellular substances are stained pink
Covering

• Applying a thin glass coverslips to protect the section

Light microscope
Different Parts of Microscope

•Eyepiece Lens
•Tube
•Arm
•Base
•Illuminator
•Stage
•Revolving Nosepiece
•Objective Lenses
•Condenser Lens
•Coarse & fine Focus
•Diaphragm
Ontogenesis of tissues

• origin of tissues and their evolution in phylogeny

• tissues cannot exist independently and in isolation, but only

integrated in hierarchically higher formations- organs and
system of organs in mandatory and continuous relationship
and interdependence.
CELL DIFFERENTIATION

• Differentiation is expressed in the division of functions between

different groups of cells
• as a result: uniform formations of specialized cells and intercellular
substance develop, providing in a community the most general
functions of the whole organism.
• That functions are:
1. barrier function – relationships between organism and external
environment
2. ensuring the relative stability of the internal environment
3. the reception of stimuli from the external environment and their
conduction of excitatory processes in the body
4. locomotor function
Tissue classification

• There are 4 basic types of tissue:

• connective tissue,
• epithelial tissue,
• muscle tissue, and
• nervous tissue.
COMMON PROPERTIES OF TISSUES

1. Regeneration
2. Hyperplasia and Hypertrophy
3. Atrophy
4. Metaplasia
5. Neoplasia
REGENERATION

• Physiological regeneration
• It occurs continuously in some tissues, the cellular elements of which
are systematically and periodically renewed.

• For example, new ones are constantly being formed in the bone
marrow to exchange old and dead erythrocytes. Epidermal cells and
epithelial cells of the intestinal mucosa are also constantly renewed.
REGENERATION

• Pathological regeneration
• It is characterized by the restoration of tissue defects due to
inflammatory and dystrophic processes, necrosis, trauma and more.

• The process begins with the growth of cells from the tissue that
borders the defect and is healthy. The growth of these cells leads to
the formation of new, young granulation tissue that fills the defect.
Hyperplasia and Hypertrophy

• Hyperplasia
• Hyperplasia is increased cell production in a normal tissue or organ.
Hyperplasia may be a sign of abnormal or precancerous changes. This
is called pathologic hyperplasia.

• It can also be due to the growth of completely normal cells. This is

called physiologic hyperplasia.
Hyperplasia and Hypertrophy

• Hypertrophy:

• Enlargement or overgrowth of an organ or part of the body due to

the increased size of the constituent cells.
Atrophy

• Atrophy is a decrease in size of a body part, cell, organ, or other

tissue. The term implies that the atrophied part was of a size normal
for the individual, considering age and circumstance, prior to the
diminution. In atrophy of an organ or body part, there may be a
reduction in the number or in the size of the component cells, or in
both.

• Certain cells and organs normally undergo atrophy at certain ages or

under certain physiologic circumstances. In the human embryo, for
example, a number of structures are transient and at birth have
already undergone atrophy.
Metaplasia

• Metaplasia is the replacement of one differentiated somatic cell type

with another differentiated somatic cell type in the same tissue.

• Typically, metaplasia is triggered by environmental stimuli, which may

act in concert with the deleterious effects of microorganisms and
inflammation.
Neoplasia

• Neoplasia is new, uncontrolled growth of cells that is not under

physiologic control. A "tumor" or "mass lesion" is simply a "growth"
or "enlargement" which may not be neoplastic (such as a granuloma).
The term "cancer" implies malignancy, but neoplasms can be sub
classified as either benign or malignant.

• There is no single mechanism by which a neoplasm arises. Many

different mechanisms give rise to neoplasms, and that is what makes
diagnosis and treatment so challenging.