Three-dimensional microscopy using a confocal laser scanning microscope

Abstract

In a scanning laser microscope detecting fluorescent light from the specimen, the depth-discriminating property of confocal scanning has been used to carry out optical slicing of a thick specimen. The recorded digital images constitute a three-dimensional raster covering a volume of the specimen. The specimen has been visualized in stereo and rotation by making look-through projections of the digital data in different directions. The contrast of the pictures has been enhanced by generating the gradient volume. This permits display of the border surfaces between regions instead of the regions themselves.