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 Introduction

 Purpose

 Effect of Fixation

 Summary
Preparation of tissue for microscopic examination ,
through a series of namely fixation , clearing,
embedding ,cutting & Staining.

Tissue decompose when removed from the body or it is

cut off from the body supply – Autolysis or
Putrefaction
 A physical or chemical process that stops as
soon as possible vital cellular processes,
maintaining with minimal alternation the
shape, volume spatial and molecular relation
between elements.

 Fixation is the foundation for the subsequent

stages in the preparation of the sections.
1.To prevent autolysis , bacterial decomposition
& putrefaction.
2. To coagulate the tissue as prevent loss of
easily diffusible substances.
3. Indurate tissues to enhance working with
glass slides.
4. To facilitate differential staining with dyes &
other reagents.
1. Inhibitions of autolysis & putrefaction
2. Preservation
3. Hardening
4. Solidification of colloid material
5. Optical differentiation
6. Effect on staining
I. Immersion

II. Perfusion

III. Perfusion – immersion

IV. Heat fixation

 It is clear that fixation is normally carried out
as soon as possible to prevent autolysis and
to reduce possible infectivity.

 Formalin is used for all routine surgical

pathology and autopsy tissues when an H & E
slides is to be produced.