Ethanol (ethyl alcohol) has long been a standard reagent used
in preparing tissues for light and electron microscopy. After
fixation, tissues are usually dehydrated with ethanol before
being embedded in paraffin or plastic. In this study we show
that the ethanol-infiltrated tissue can be frozen and sectioned
directly, without embedding. When tissue impregnated with ethanol
is cooled below about -117¡C with liquid nitrogen, the ethanol
solidifies without appreciable crystallization. The frozen tissue
can then be sectioned in a commercial cryoultramicrotome that
is set at -155 to -170°C to produce semithin frozen sections
(0.5-3 μm thick) for light microscopy, or ultrathin frozen sections
(50-100 nm thick) for electron microscopy. Sections are picked
up and mounted on glass slides or EM grids by means that are
in current use for ice ultrathin frozen sectioning. Since there
is no apparent freezing damage, the morphology in these ethanol
frozen sections of unembedded tissue appears generally quite
good, often resembling that obtained by conventional EM techniques.
Examples are provided that illustrate the use of this material
for immunocytochemistry at the light and electron microscope
levels.