SPI-Mark™ Colloidal Gold Reagents
Methods of Etching
(A) For sections of tissue post fixed with osmium tetroxide, treat sections with
1-4% hydrogen peroxide solution for 5-10 min at room temperature. This is an
oxidising reaction which removes reduced osmium metal and makes the
sections more hydrophilic. Wash sections thoroughly in water before
performing normal incubations.
(B) Treat sections of osmium post fixed tissue with saturated sodium
metaperiodate (0.1% diluted in distilled water) for 3-5 min at room
temperature. This acts in a similar way to hydrogen peroxide (a).
(C) Treat epoxy sections with a saturated solution of sodium hydroxide in
ethanol or methanol for 30 min at room temperature. This removes epoxy resin
from sections. Difficult to control for EM sections.
(D) Treat acrylic sections with 70-100% ethanol. This partially dissolves the
methacrylate bonds and removes resin slowly. Acrylics feel soapy when
treated with alcohol for this reason.
(E) Treat acrylic resins with weak hydrochloric acid (0.01-0.1%) for 10 min (on
gold grids only). This gradually removes the resin by interfering with the ester
bonds. Difficult to control without affecting antigenicity.
(F) Treat sections with 0.1% trypsin in TBS for 30 min at 37°C. This unmasks
antigens from un-reactive precursors and digests surrounding epitopes. Useful
for treating heavily fixed tissue (e.g. after glutaraldehyde). Difficult to control
selective digestion without interfering with specific epitopes.
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Thursday May 17, 2012
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