Abstract
Biological material is organized in four dimensions: three spatial ones and a temporal one. Light microscopy is able to visualize biological objects in their natural watery condition and during their temporal development. Improved imaging is the optical sectioning property by which the contributions from out-of-focus areas in the specimen are effectively suppressed. In normal microscopy, these contributions lead to a strong reduction in the available image contrast. A three-dimensional microscope is obtained where each data point as collected represents the quantity of the specific contrast parameter used at a certain point in space. Deconvolution techniques have been developed for eliminating the out-of-focus information from conventional fluorescence microscopy. Confocal microscopy can deliver directly clear optical sections without the use of time-consuming image reconstruction algorithms. Image processing can also be used to enhance the confocal images. Computer-generated stereoscopic images are also used for the visualization of the three-dimensional biological information. This chapter discusses the number of aspects of confocal imaging, especially the advantages and drawbacks of the various scanning approaches.
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