Confocal microscopy: a powerful technique for biological research

Abstract

Confocal microscope permits the generation of three-dimensional images of biological and nonbiological specimens. The efficacy of this technique lies in the elimination of out-of-focus glare by spatial filtering, utilizing a point source of light for excitation, and a pinhole confocal with the excitation pinhole in front of the detector. A combination of transverse resolution with noninvasive optical sectioning results in very high quality images of biological specimens. Several combination of lasers can be coupled to the fibre optics of the scanning unit in order to increase the number of excitation wavelengths. Powerful softwares that display and analyse 3-D data are currently available. Laser scanning confocal microscopy has proved to be most suitable for the analysis of structural details of thick specimens and promises to be of great potential in providing 3-D volume renderings of living cells and tissues over time.