Microscopy: Light and Confocal

Authored by: Casey R. Lu , Peter B. Kaufman , Leland J. Cseke

Handbook of Molecular and Cellular Methods in Biology and Medicine

Print publication date:  December  2011
Online publication date:  April  2016

Print ISBN: 9781420069389
eBook ISBN: 9781439881958
Adobe ISBN:

10.1201/b11351-45

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Abstract

Major conceptual breakthroughs have resulted and continue to be discovered in cellular and molecular biology via the techniques of light microscopy (LM) and confocal microscopy (CM). For investigators conducting studies on in situ localization of specific mRNAs, reporter gene expression, protein and ion localization in animal or plant tissues and organs, LM and CM protocols are required. In addition to in situ localization, CM allows researchers to generate three-dimensional (3-D) optical sections of living tissues and cells using computer programs for data and section analyses, fluorescent markers, transgenic lines (e.g., containing green fluorescent protein [GFP]), and filter systems that allow investigators to observe fluorescence images of specific proteins, nucleic acids, and ions within whole tissues or organs. Multiphoton microscopy is a newer technique that allows the same kind of 3-D optical sections that one may produce with a confocal microscope, but without the problems of photobleaching, and it allows for deeper penetration due to the longer wavelengths used in this microscope.

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