Abstract

Confocal scanning microscopes are known to possess superior optical sectioning capabilities compared to conventional microscopes. Out-of-focus contributions in a through-focus series of images are significantly reduced by the confocal geometry but not completely removed. This paper reports our initial investigations Into a posterioriimage processing (i.e., deconvolution) for further improvement of depth resolution in confocal microscopy. This project is part of a larger effort In laser scanning fluorescence microscopy for biological and biophysical analyses in living cells. The instrument is built around a standard inverted microscope stand, enabling the use of standard optics, micromanipulation apparatus, and conventional (including video) microscopy in conjunction with laser scanning. The beam is scanned across the specimen by a pair of galvanometer-mounted mirrors driven by a programmable controller which can operate In three modes: full raster scan; region of interest; and random-access (point-hopping). After taking a scout image with laser scanning or video, the user will select isolated points or regions of interest for further analysis via a graphic user interface implemented on the system’s host computer. Experimental parameters such as detector integration times are set up with a window-style menu.

© 1988 Optical Society of America