Embedding in Resins
for Immunocytochemistry
Biological specimens
are embedded in resin so that they can be thin sectioned. Many different
resin formulas are currently available and all have been used for post-embedding
labeling (i.e. where antibodies are applied to thin sections of the tissue)
with variable results. Specialized resin formulas have been developed
specifically for use in immunocytochemistry. These include the Lowicryl
resins, LR White and LR Gold, Unicryl and MonoStep. They all have low
viscosity and can be polymerized, at low temperatures, with ultra violet
(UV) light.
The most common embedding
protocol for these resins is called the progressive lowering of temperature
(PLT) method where the sample is progressively cooled during the dehydration
steps. The specimens are then transferred to resin and the blocks are
polymerized, at low temperature, by exposure to UV light.
An alternative method
for embedding specimens in these resins is by freeze
substitution. This method consists of freezing the specimen and then
substituting the frozen water in the specimen with a liquid solvent. Follow
the following links for more details on the theory of
freeze substitution and for a simple protocol of
freeze substitution.
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