Introduction to Histopathology

Inam ullah
Lecturer MLT (SIAHS)
 Introduction
• Histology is the microscopic study of normal
tissues while histopathology is the study of
diseased tissues.

• The techniques required for histological or

histopathological microscopic studies are termed
Histotechniques (Histological Techniques or
Microtechinques).
 Introduction
• Infact it is the study of procedures or stages
to reach the final stained slide of the
specimen for microscopic examination.
 Introduction
• The persons responsible for performing
these procedures are called
histotechnologist and the field of
specialization is termed as histotechnology.
 Introduction
• These techniques, employed in a
histopathology laboratory, help a
histopathologist to determine whether a
patient has malignancy or some other
disease process.

• The microscopic examination involves

following steps:
 Documentation
• Every specimen is given the histopathology
[HP] number which is pen down at top
corner of the request slip, in a specified
record register and on the top of container.
 Fixation
• It is a process by which the constituents of
cells are fixed in a physical and partly also
in a chemical state, so that they will
withstand subsequent treatment with
various reagents with a minimum of loss, or
decomposition.
• This is achieved by exposing the tissue to
chemical compounds called fixatives.
 Gross Examination
• The specimen is examined grossly by noting
anatomical features of it.

• Representative sections are selected for further

procedures while small biopsies are selected as
a whole.
 Decalcification
• In case of calcified tissue (bone), calcium
salts are removed from the bony tissue.

• It is done before gross examination.

 Tissue Processing
• It involves a number of stages in which
selected portions of tissue are passed
through a number of chemicals in a
sequence.
 Tissue Processing
• In this stage tissues are impregnated with a
solidifying medium to facilitate its fine
sectioning.
 Tissue Processing
• Following four stages are involved:

i. Completion of Fixation
ii. Dehydration (ethyle alcohol)
iii. Dealcoholization or Clearing(xylin)
iv. Impregnation or Infiltration
 Embedding
• In this step all processed tissues are blocked
out in a solidifying medium (,.e.g. paraffin
wax) which is used in the impregnation
stage.
 Sectioning
• Paraffin blocks of tissues are then sectioned by
means of a device termed as Microtome.

• The sections which are produced are capable of

transmitting light and are at a micron level
thickness.
 Staining
• Tissue sections are picked onto slides and
staining is performed in order to examine
different tissue components under the
microscope.
 Mounting
• Cover slip is applied on each section using
appropriate medium to obtain permanent
preparation for microscope