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During our combined 35 plus years of operating confocal microscopes and managing core microscopy facilities, and through teaching our Basic Confocal Microscopy Workshop at several venues, we have found that students and technicians who are novice users of confocal microscopes are often instructed by their mentors to go to the confocal system and collect some images. Often the implied message is that...
To harness the power of fluorescence for biological microscopic imaging, a number of additional components must be introduced to the standard light microscope. The most important modifications are (1) a strong illumination system that provides suitable wavelengths for exciting a specific fluorochrome, (2) some mechanism to limit the illumination beam to specific wavelengths so that only the fluorochrome...
A key difference between confocal microscopy and widefield microscopy is that the aim of confocal is to explore the structure and structural relationships along the optical (Z) axis as well as in the X-Y plane. In other words, the investigation of spatial relationships is being evaluated in at least three dimensions. Often, this involves the collection of a series of planar images along the Z axis...
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