rev:May 27, 1997
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BIOBOND (tm) & BIOMOUNT(tm)
cat#RDI-BB20 $112.00/20ml vial $94.00/vial 3 or more
For light microscopy and electron microscopy the choice of specimen preparation
si critical for the preservation of antigens in the sample. Of greatest
importance in the preparation schedules are the specimen fixation and embedding.
this 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 maximize the
specific signal and minimize 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 minimize this possibility by coating the surface with a material that
is of low reactivity towards reagents.
A new special slide coating solution, BIOBOND (tm) is available which produces
a very strong adhesion between the glass and the tissue section for subsequent
incubation. Biobond (tm) coats the slide with a protective layer to minimize
interaction of charged glass surface with reagent. This is also of particular
importance for reproducibility of results because of the variations that
occur between glass slides obtained from different sources in different
countries. It is particularly effective for use with severe incubating conditions
such as those used in in situ hybridization. Biobond (tm) is suitable for
all kinds of tissue specimens including paraffin wax or resin sections, cell
smear, cytospins or cryostat sections. Biobond (tm) is supplied in 20ml unit
volumes, sufficient to coat at least 1000 slides.
Simple Coating Procedure:
1) Make a 2% solution of Biobond (tm) is acetone (eg 2ml in 100ml acetone).
Use fresh solution each time.
2) Place slides in a plastic or metal holder and immerse in Biobond solutions
for 4 minutes.
3) Rinse the slides in clean water for 5 minutes
4) Air Dry
-The slides are then ready for mounting sections. They may be stored indefinitely.
BIOMOUNT (tm) TISSUE SECTION MOUNTING MEDIUM
cat#RDI-BM100 $94.00/100ml vial $62.00/vial 3 or more
Biomount (tm) reduces fading of immunogold/silver signals in sections on
glass slides. It is suitable for both resin and wax embedded tissue
sections.
Some mounting media oxidize rapidly on exposure to air, forming carboxyl
groups. This may be especially so when sections have been cleared in a solution
containing aldehyde groups. It is sometimes observed that the immunogold/silver
stain fades after a few weeks or even in a shorter time, from sections that
have been mounted with these media under cover slips. The silver is still
present but has formed translucent silver carboxylate salts. Visibility can
be retrieved by removing the cover slip and washing in xylene (xylol) and
then immersing the slide in photographic developer, but this is a tedious
procedure. Biomount (tm) prevents this fading because of its low oxidizing
properties.
Biomount (tm) is miscible with xylene and may be applied to sections on slides
in the normal manner following dehydration and immersion in xylene. Labelling
will retain its intensity and contrast following the use of Biomount (tm).
Sections that have been counterstained with normal stains will also retain
their color intensity.
RDI Division of Fitzgerald Industries Intl
34 Junction Square Drive
Concord MA 01742-3049
USA
phone (978) 371-6446 or (800) 370-2222
fax (978) 371-2266
EMAIL:antibodies@fitzgerald-fii.com