Five Major Steps of Paraffin Embedding | Immunostaining

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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 par
affin. Immunostaining provides the basic guide
for fixation, microtome sectioning and staining
of paraffin embedded tissue sections.
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Five Major Steps that Involved In Paraffin
Embedding
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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.
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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?.
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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.
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To know more about paraffin embedding, visit at
http//immunostaining.info/ihc-principle/paraffin-
embedding/
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