Imaging, representation and analysis of 3-dimensional biological structures by confocal microscopy

Abstract

Confocal scanning laser microscopy is a new technique capable of visualizing the three-dimensional structure in suitable objects. As further explained in fig. 1 the basic aspect of this type of microscopy is that one and the same object point in the specimen is both optimally illuminated from a laser illuminated pinhole as well as optimally imaged on a detector pinhole. Due to the specific properties of this optical arrangement not only the lateral resolution is substantially improved, but also an excellent optical sectioning property comes available. The latter means that only information from a chosen plane in the specimen will contribute to the image, while the influence of the layers above and below is suppressed. The important advantage over elec- tronmicroscopy, apart from the speed of the technique, is that to make the 3-D information accessible in a specimen no mechanical sectioning of the material is necessary.