Confocal  microscopy is a very powerful tool and has many advantages over wide-field  fluorescent  microscopy. One of these advantages is the ability to look at thin optical sections from a sample of cells or tissue without physically cutting the sample. This enables the user to get rid of the out of focus light associated with wide-field fluorescent microscopy.

Image diagram provided by  Les.Boland

The above diagram  represents the optical path on a laser point scanning confocal microscope.A small diaphragm, situated in a conjugated focal plane, prevents out of focus light reaching the detected. The diameter of this diaphragm controls the depth of the optical section.Images captured will have improved Z resolution and fine detail that is normally obscured by the out of focus blur assosiated with wide-field fluorescent microscopy will be seen.


Z sectioning allows the optically collection of  images through the depth of the sample. This series of images can then be reconstructed into a high resolution 3D image.

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Mitotic cells  in 3D and Gallery showing  optical Z stack. L.Black R11




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Last modified: 02/25/16