REAL-TIME HIGH-PERFORMANCE CONFOCAL LASER SCANNING OPTICAL MICROSCOPE

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A no-moving-parts, 30-frames-per-second, laser beam scanning confocal reflected light and fluorescence microscope has been developed. Acousto-optic beam deflectors, controlled by all-digital electronics, move a laser beam in a 512-line, interlaced raster. The light enters an inverted microscope through the rear epifluorescence light port and is imaged at the object by the microscope objective. Reflected or fluorescent light returns through the objective, exits through the camera port, and is imaged onto the photocathode of an image dissector tube (IDT). Confocality is provided by raster-scanning the IDT aperture coincident with the congruent image of the laser beam incident on the object. Real-time, reflected, and fluorescent light images of a variety of biological objects have been produced. Improved electronics and simplified optics overcome the usual dispersive problem with acousto-optics to enable the laser wavelength to be easily changed, resulting in a much more flexible system.