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Multi-Photon Excitation Microscopy: An Old Idea in Quantum Theory Applied to Modern Scientific Problems

Published online by Cambridge University Press:  02 July 2020

David W. Piston
David W. Piston*
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN37232.
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Extract

Multi-photon excitation microscopy provides attractive advantages over confocal microscopy for three-dimensionalry resolved fluorescence imaging and photochemistry. The most commonly used type of multi-photon excitation is two-photon excitation where simultaneous absorption of two photons leads to a single quantitized event. The powerful advantages of using two-photon excitation microscopy arise from the basic physical principle that the absorption depends on the square of the excitation intensity. In practice, two-photon excitation is generated by focusing a single pulsed laser through the microscope. As the laser beam is focused, the photons become more crowded, but the only place at which they are crowded enough to generate an appreciable amount of two-photon excitation is at the focus. Above and below the focus, the photon density is not high enough for two of them to interact with a single fluorophore at the same time. This dramatic difference between confocal and two-photon excitation microscopy is shown in Fig. 1.

Type
Multi-Photon Excitation Microscopy
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 1180 - 1181
Copyright
Copyright © Microscopy Society of America

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References

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8 Support for this work comes from NIH (DK53434), NSF (DBI-9871063), The Vanderbilt Diabetes Research and Training Center (DK20593), and The Vanderbilt-Ingram Cancer Center (CA68485).Google Scholar