INTRODUCTION TO

HISTOLOGY


Fernando J. Peraldo, M.D., MPH
HISTOLOGY
is derived from the Greek word for a tissue
"Histos", and "-logos" = the study of cells and
the extracellular matrix of tissues
"Microscopic Anatomy - includes
understanding of the structure and function of
cells, tissues, organs and organ systems
the study involves the use of microscopes
(light and electron) as basic tools.
HISTOLOGY
the body can be seen to be formed of different levels
of organization, with increasing levels of complexity
LEVELS OF ORGANIZATION
Cells
Tissues
Organs
Organ Systems
Organism

each of which plays important roles in the physiological
homeostasis of the body.



LEVELS OF ORGANIZATION

THE CELL - defined as the smallest basic
structure of higher organisms capable of
independent existence
TISSUES - are groups of cells of similar
function and origin that form functional units
ORGANS - represent an even greater measure
of complexity and are composed of various
tissues
ORGAN SYSTEM - composed of several organs

BASIC TECHNIQUES: Preparation of
histological sections

1. Fixation
- to preserve tissues and prevent structural
change or breakdown of the components of the
tissues
- needs to preserve the tissues as close as
possible to the living state
- the fixatives commonly stabilize or denature
proteins
Ex. Formaldehyde - cheap and penetrates
tissues rapidly


BASIC TECHNIQUES: Preparation of
histological sections

2. Embedding
- thin sections require tissues to be infiltrated after fixation with
embedding substances that imparts rigid consistency to the tissue
- the most commonly used embedding or support medium is
paraffin wax and plastic resins
STEPS:
Dehydration - remove all the water from the tissue
- achieved using an ascending series of alcohols
(70%, 95%, 100%)
Clearing - tissue immersion in a wax solvent such as xylene or
chloroform
- the tissue is then transferred to molten paraffin wax (in an
embedding oven) for a couple of hours


BASIC TECHNIQUES: Preparation of
histological sections

3. Microtomy
- sections of the tissue embedded in the wax
block are cut on a machine, known as a microtome,
using special knives
- series or ribbons of sections are cut at a thickness of
6-8mm.
- the sections are transferred to the surface of a hot
waterbath then collected on glass microscope slides
(standard dimensions of 3 x 1 inches)
- In order for the sections to adhere to the slides they
are dried for up to 24 hours in a drying oven


BASIC TECHNIQUES: Preparation of
histological sections
4. Staining
- the most common staining technique is known as
Hematoxylin and Eosin (or H&E) staining
STEPS:
- remove wax using a wax solvent such as xylene
- hydrate the slide using a series of descending
alcohols (100%, 95%, 70%) and then water
- immerse the slide in Hematoxylin stain, rinsed in
running water (preferably alkaline), followed by
staining with Eosin, and rinsing in water.


BASIC TECHNIQUES: Preparation of
histological sections

5. Permanent Mounting
- After staining the sections are again
dehydrated with ascending alcohols (95%,
100%) and xylene, prior to covering with a
mountant and a glass coverlip
- the slide is left for at least 24 hours for the
mountant to dry
- the finished (permanent) slide with its stained
tissues can then be examined under the
microscope.


BASIC TECHNIQUES: Preparation of
histological sections

Frozen sections
- A rapid alternative to wax embedding
- use of cryostat (a microtome operated in a low
temperature cabinet,
usually about -30 C, then be stained and mounted
in a suitable water-soluble mountant.
Total preparations
- Use in a very thin membrane. The tissue does not
need cutting on a microtome, but can be stained,
mounted and examined directly. Not as 2-
dimensional as histological sections, and adjustment
of focus is necessary during examination.


BASIC TECHNIQUES: Preparation of
histological sections

Cell Smears
- a form of histological preparation that does
not require sectioning
Example: blood or bone marrow smears
swabs or scrapings of epithelial
cells (e.g. from the oral cavity,
cervix uteri).


Histological classification of animal tissues

Four basic types of tissues
muscle tissue
nervous tissue
connective tissue
epithelial tissue
All tissue types are subtypes of these four
basic
tissue types (for example, blood cells are
classified as connective tissue, since they
generally originate inside bone marrow).
SPECIAL TERMS
Epithelium: the lining of glands, bowel, skin and some
organs like the liver, lung, kidney
Endothelium: the lining of blood and lymphatic
vessels
Mesothelium: the lining of pleural and pericardial
spaces
Mesenchyme: the cells filling the spaces between the
organs, including fat, muscle, bone, cartilage, and
tendon cells
Blood cells: the red and white blood cells, including
those found in lymph nodes and spleen

Neurons: any of the conducting cells of the nervous
system
Germ cells: reproductive cells (spermatozoa in
men, oocytes in women)
Placenta: an organ characteristic of true mammals
during pregnancy, joining mother and offspring,
providing endocrine secretion and selective exchange
of soluble, but not particulate, blood-borne
substances through an apposition of uterine and
trophoblastic vascularized parts
Stem cells: cells able to turn into one or several of the
above types

END OF LECTURE
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