Serial Sectioning Of Cells In Three Dimensions With Confocal Scanning Laser Fluorescence Microscopy (Fl-CSLM): Microtomoscopy

The discrimination of out of focus contributions in fluorescence microscopy possible in a confocal setup will establish itself as a supplement to conventional fluorescence microscopy. The improvement of the contrast compared with conventional fluorescence microscopy depends mainly on the density of the fluorescing material and the thickness of the sample. The term thickness, that which microscopists refer to as the size of the specimen along the optical axis, will gain a new quality since a confocal fluorescence microscope may reveal totally different features when recording data in planes that are 0.3μm apart. Differences that have in the past been neglected suddenly become important. The following article will outline important features in the application of confocal fluorescence microscopy in the biological sciences, point out its limitatk'ns, and draw attention to expected developments.