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Optical Sectioning in Fluorescence Microscopy by Confocal and 2-Photon Molecular Excitation Techniques

Published online by Cambridge University Press:  14 March 2018

M.B. Cannell  and
C. Soeller
M.B. Cannell
Affiliation:
St. George's Hospital Medical School, Cranmer Terrace, LondonSW17 ORE
C. Soeller
Affiliation:
St. George's Hospital Medical School, Cranmer Terrace, LondonSW17 ORE

Extract

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Fluorescence microscopy has proved to be an invaluable tool for biomedical science since it is possible to visualise small quantities of labeled materials (such as intracellular ions and proteins) in both fixed and living cells, However, the conventional wide field fluorescence microscope suffers from the disadvantage that objects outside the focal plane also fluoresce (in response to the excitation light) and this leads to a marked loss of contrast for objects in the focal plane, This is especially a problem when the fluorescent probe is distributed throughout the thickness of the cell and the cell is thicker than about 1 µm. The confocal microscope overcomes this problem by illuminating the preparation with a point source of excitation light and limiting the collection of light with a pinhole that is confocal with the illumination source.

Type
Research Article
Information
Microscopy Today , Volume 5 , Issue 8 , October 1997 , pp. 12 - 15
Copyright
Copyright © Microscopy Society of America 1997

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