Abstract
Confocal microscopy was developed in 1957 by Minski, who was awarded a patent for this work in 1961. Since that time many advances have been in new designs and implementations. There are two basic classes of confocal microscope: the Nipkow-disk-based confocal microscope which allow real-time direct viewing of the sample, and the confocal scanning laser microscopes (CSLM). The CSLM will be focussed on in this presentation.The CSLM scans a focussed beam over, across or through the sample, collecting the reflected, scattered or emitted light. This light is directed towards an optical spatial filter, which passes light returning from the on-axis focus position, and rejects light that is returning from anywhere else. By detecting the light passing through the spatial filter synchronously with the moving beam and/or sample, an image that has only in-focus information is acquired. Typically a set of images is acquired as the sample is moved through focus: from this “image stack” both an extended-focus image and topographic information can be obtained.
Publisher
Cambridge University Press (CUP)