We report on the development of a laser scanning fluorescence microscope possessing several features which facilitate its application to biological and biophysical analyses in living cells. It 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. A full 512x512 pixel image can be acquired in one second. In region of interest mode, several subareas of the field can be selected for more rapid or detailed analysis. For those cases where the time scale of the observed phenomenon precludes full-field imaging, or where a full-field image is unnecessary, the random access mode enables an arbitrary pattern of isolated points to be selected and rapidly sequenced through. Via a graphical user interface implemented on the system's host computer, a user will be able to take a scout image either with video or a full-field laser scan, select regions or points on the scout image with a mouse, and set up experimental parameters such as detector integration times with a window-style menu. The instrument is designed to be a flexible testbed for investigating new techniques, without compromising its utility as a tool for biological research.© (1988) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.