Abstract

In recent years, the ability to image morphological dynamics and physiological changes in living cells and tissues has been greatly advanced by the advent of scanning laser confocal microscopy. Confocal microscopes employ optical systems in which both the condenser and objective lenses are focused onto a single volume element of the specimen. In practice, galvanometer-driven mirrors or acousto-optical deflectors are used to scan a laser beam over the specimen in a raster-like fashion through an epifluorescence microscope. The incident laser beam, as well as the collected fluorescent light, are passed through pinhole or slit apertures in image planes that are conjugate to the plane of the specimen. This method of illumination and detection prevents fluorescent light which is generated above and below the plane-of-focus from impinging on the imaging system's photodetector, thus rejecting much of the fluorescent light which normally blurs the image of a three-dimensional fluorescent specimen.

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