Paraffin Embedding

Paraffin embedding is a method of tissue

preservation for immunohistochemistry. When we
generating paraffin embedded tissue sample,
the tissue should be fixed
before embedding in paraffin. Immunostaining
provides the basic guide for fixation, microtome
sectioning and staining of paraffin – embedded
tissue sections.
Five Major Steps that Involved In Paraffin Embedding

Fixation Dehydration

Transparentizing Immersion

Embedding
1. Fixation
This is the process by which the constituents of cells and
tissue are fixed in a physical and chemical state so that
they will with stand subsequent treatment with various
reagents with minimum loss of architecture. This is
achieved by exposing the tissue of chemical compound,
call fixatives.
2. Dehydration
Dehydration removes water completely, creates a
condition for the next step and hardens the tissue of
interest.
3. Transparentizing
After dehydration, the tissue of interest requires a
transparentizing step because the dehydrating agent
used in the previous step is immiscible with the paraffin
from one of the subsequent steps. The addition of
transparent reagent helps paraffin absorb into the tissue.

4. Immersion
After transparentizing, the tissue can be immersed in
molten paraffin wax so that it adsorbs the wax-
substituting transparent agent. Based upon the
melting point of wax, immersion should be performed
at 54-64℃.
5. Embedding
This is a process of treating the tissue in a paraffin box
so that the paraffin wax cools down and solidifies. The
treatment conditions (using ethanol and xylene as an
example) are shown in the table below. After cooling is
completed, the tissue will be ready for sectioning and
suitable for storage.
 To know more about paraffin
embedding, visit at
http://immunostaining.info/ihc-
principle/paraffin-embedding/
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