| Summary: | SUMMARY Acrolein is a potent fixative that provides both excellent preservation of ultrastructural
morphology and retention of antigenicity, thus it is frequently used for immunocytochemical
detection of antigens at the electron microscopic level. However, acrolein is not
commonly used for fluorescence microscopy because of concerns about possible autofluorescence
and destruction of the luminosity of fluorescent dyes. Here we describe a simple protocol
that allows fine visualization of two fluorescent markers in 40-mm sections from acroleinperfused
rat brain. Autofluorescence was removed by pretreatment with 1% sodium borohydride
for 30 min, and subsequent incubation in a 50% ethanol solution containing 0.3%
hydrogen peroxide enhanced fluorescence labeling. Thus, fluorescence labeling can be used
for high-quality detection of markers in tissue perfused with acrolein. Furthermore, adjacent
acrolein-fixed sections from a single experiment can be processed to produce high-quality
results for electron microscopy or fluorescence labeling.
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