Three-dimensional visualization of Golgi-stained neurons by a projection X-ray microscope converted from a scanning electron microscope.

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Abstract
In trying to obtain the 3-demensional images of [rat] neuronal elements, Golgi-stained thick sections were examined by a projection X-ray microscope converted from a scanning electron microscope. An attachment unit for the projection X-ray microscope consists of a target holder, specimen stage and film cassette. Two kinds of target material, Ti and Au, were tested. The system was operated at 10-30 kV. The electron beam into the target was .apprx. 20 nA with the use of a pointed cathode. The stereopair of X-ray micrographs were taken by tilting the specimens. Quality and contrast of the X-ray micrograph taken with a Ti target was better than that with a Au target. The spatial resolution was estimated .apprx. 1-2 .mu.m for 100-350 .mu.m thick sections with a Ti target. Compared with the light microscope, the strong penetrating ability of X-ray permitted visualization of the internal structures of optically opaque materials. Almost all objects were in focus in X-ray micrographs even for relatively thick sections (100-350 .mu.m). the 3-dimensional morphology of neurons and glial cells was demonstrated by a stereopair of X-ray micrographs.

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