An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy.

It is found that confocal imaging gives greatly enhanced images of biological structures viewed with epifluorescence, and the improvements are such that it is possible to optically section thick specimens with little degradation in the image quality of interior sections.

Abstract

Scanning confocal microscopes offer improved rejection of out-of-focus noise and greater resolution than conventional imaging. In such a microscope, the imaging and condenser lenses are identical and confocal. These two lenses are replaced by a single lens when epi-illumination is used, making confocal imaging particularly applicable to incident light microscopy. We describe the results we have obtained with a confocal system in which scanning is performed by moving the light beam, rather than the stage. This system is considerably faster than the scanned stage microscope and is easy to use. We have found that confocal imaging gives greatly enhanced images of biological structures viewed with epifluorescence. The improvements are such that it is possible to optically section thick specimens with little degradation in the image quality of interior sections.