For light microscopy and electron microscopy, the choice of specimen
preparation is critical for the preservation of antigens in the sample. Of
greatest importance in the preparation schedules are the specimen fixation
and embedding. The protocol must satisfy the requirements for preservation
of structural integrity and antigenicity.
Having prepared the tissue specimen for immunolabelling, it is then
imperative to perform the incubations with a protocol designed to maximise
the specific signal and minimise the background. Some incubation conditions
may cause tissue sections to be removed from the glass slide. Typical tissue
section adhesives such as poly-L-lysine, Elmer's glue, chrome alum, etc are
not suitable for use with immunogold labelling because of the increased
background caused by attraction of gold particles to the adhesive on the slide.
In addition, the surface of glass slides is uneven and is activated by the
silicon tetrahedral structure. This provides active sites for absorption of
proteins or reactions with chemicals and reagents. It is therefore important
to minimise this possibility by coating the surface with a material that is
of low reactivity towards reagents.
SPI Supplies now offers a special slide coating solution, BioBond™, which
produces a very strong adhesion between the glass and the tissue section for
subsequent incubations. BioBond coats the slide with a protective layer to
minimise interaction of charged glass surface with reagents. This is also of
particular importance for reproducibility of results because of the
variations that occur between glass slides obtained from different sources
and in different countries. It is particularly effective for use with severe
incubating conditions such as those used in
In-situ hybridization.
BioBond is suitable for all kinds of tissue specimens including paraffin
wax or resin sections, cell smears, cytospins or cryostat sections.
A 2% solution is used for the coating of slides.
BioBond is supplied in 250 ml unit volumes, sufficient to coat at least 1000
slides.
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