Abstract
To obtain stereographic images of tissues containing both free and sectioned surfaces, backscattered electron (BS) imaging was tested with both resin and de-embedded specimens. Although the ultrastructure of several organelles in cells could be identified on a sectioned plane of the resin-embedded tissues, threedimensional images of tissues were difficult to obtain. On the other hand, threedimensional images showing both the free and sectioned surfaces of tissues could be obtained by scanning electron microscopic observation of the de-embedded specimens stained with heavy metals in BS mode. Organelles in the sectioned plane could be recognized with enough contrast to discriminate the different cellular components even after the de-embedding process. Our present method would make it easier to obtain images of rare structures barely detectable by the conventional cracking methods and will contribute to a three-dimensional analysis of biological specimens.
