SPI-Mark™ Colloidal Gold Reagents
Instructions for Use and Sample Protocols
Silver Enhancing
Silver enhancing may be used for EM studies where the gold particles are too
small to be seen in the available microscope or to grow them from any original
size to a size that gives a more comprehensive view of labeling at lower
magnifications. The method is quite easy to perform for any original gold
particle size chosen. In order to ensure that the gold labels are not removed
from the section it is common to fix the section for 10 minutes in 1%
glutaraldehyde in water before silver enhancing. Using the
Silver Enhancing Kit for LM and EM
(SEKL) a drop of enhancer and initiator are
mixed together on a clean petri dish and the gold labelled EM section placed
on the surface or even immersed in the droplet. The time of enhancement must
be judged empirically but approximately 5-10 minutes is usually sufficient to
grow from 1nm to 5nm, or from 5nm to 20nm. It is temperature dependent.
The enhancement process is stopped by washing in distilled water and the
grids may be re-enhanced after examining in the EM until the chosen size is
achieved. Provided the sections are not exposed to an electron beam, silver enhanced sections
may be further immunolabelled in a subsequent incubation
procedure. This may provide another method of double labeling.
For LM studies silver enhancing is a necessary but straightforward procedure.
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Thursday May 17, 2012
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