Histology

Histology:

•is the study of the tissues of the body and how these
tissues are arranged to constitute organs.
•Involved in all aspects of tissue biology, with the focus
on how cells’ structure and arrangement optimize
functions specific to each organ.

Tissue have two

interacting components:

•Cells
•Extracellular Matrix (ECM).
EXTRACELLULAR MATRIX

The ECM
• Supports the cells and the fluid that transports nutrients to the
cells, and carries away their catabolites and secretory products.
• Consists of many kinds of macromolecules, most of which form
complex structures, such as collagen fibrils and basement
membranes.

• The cells produce the ECM and are also influenced and sometimes controlled by
matrix molecules.
• Cells and matrix interact extensively, with many components of the matrix
recognized by and attaching to cell surface receptors.
• Many of these protein receptors span the cell membranes and connect to structural
components inside the cells.
CELL & T ISSUE • Cell culture has been invaluable in studying the functions of
CULTURE molecules.
• It allows the direct observation of cellular behavior
primary cell cultures

• In preparing cultures from a tissue or organ, cells must be dispersed mechanically or

enzymatically.
• Once isolated, the cells can be cultivated in a clear dish to which they adhere,
usually as a single layer of cells
• The ideal microscopic preparation is preserved so that the tissue on the slide has the
same structure and molecular composition as it had in the body
Fixation

•cell and tissue structure is treated and preserved immediately after removal
from the body to avoid tissue digestion by enzymes present within the cells
(autolysis) or by bacterias.
•usually involves immersion in solutions of stabilizing or cross-linking
compounds called fixatives.
Popular fixatives
Aldehydes Picrates Alcohol Oxidizing Agent

• Formalin – • Bouin’s • Ethanol • osmium tetroxide

Formaldehyde solution • Methanol • potassium
• Glutaraldehyde dichromate
• Paraformaldehy • Potassium
de (PFA) permanganate
• chromic acid
Embedding &
Sectioning
 • . Tissues are embedded in a solid medium to facilitate
sectioning.
• In order to cut very thin sections, tissues must be
Embedding
infiltrated after fixation with embedding material that
imparts a rigid consistency to the tissue.
Embedding
materials
include

•paraffin - is used routinely for

light microscopy,
•plastic resins - used for both
light and electron microscopy

Paraffin Embedding
• or tissue impregnation
• When tissue is filled with liquid paraffin, the impregnated tissue then hardens in a small
container of paraffin at room temperature.
• is preceded by two other main steps:
• Dehydration
• clearing
 • A hardened block containing tissue and paraffin is placed
in an instrument called a microtome and sliced by the
steel blade into extremely thin sections.
Sectioning
• Paraffin sections are generally cut at 1-10 μm thickness
• The very thin sections are placed on glass slides and
stained
 • Most cells and extracellular material are completely colorless,
and to be studied microscopically sections must typically be
stained (dyed).

Staining • Cell components such as nucleic acids with a net negative

charge (anionic) stain more readily with basic dyes and are
termed basophilic; cationic components, such as proteins with
many ionized amino groups, have affinity for acidic dyes and
are termed acidophilic.
Examples of
stains (dyes)

• Basic dyes - toluidine blue, alcian blue, methylene

blue, hematoxylin
• Acid dyes - eosin, orange G, and acid fuchsin
• Of all staining methods, the simple combination of
hematoxylin and eosin (H&E) is used most commonly