Applications of Confocal Microscopy to Studies of Sea Urchin Embryogenesis

Abstract

The principles, advantages, and some of the general applications of confocal microscopy are presented by Wright et al. ([I] in this volume). The primary benefit of the confocal microscope derives from its ability to obtain optical sections from which out-of-focus information has been removed by the confocal aperture. The background rejection of the confocal aperture is only one consideration, however. Specimen preparation is an equally important factor that determines whether or not sections of sufficient resolution and contrast can be successfully obtained from deep within a thick specimen (see Cheng and Summers, 1990). Once confocal images of adequate quality are collected from the interior of an entire specimen or from a limited area of interest within that sample, the optical sections are stored individually in a digital form on the host computer. Subsequently, these data sets can be transformed into three-dimensional, stereoscopic, or extended-focus views that allow an entire specimen to be viewed in focus (Boyde, 1990).

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