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Second Harmonic Imaging Microscopy: A New Non-Linear Optical Modality for Cell Membrane Physiology

Published online by Cambridge University Press:  02 July 2020

Paul Campagnola ,
Aaron Lewis  and
Leslie M. Loew
Paul Campagnola
Affiliation:
Department of Physiology and Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, CT , 06030
Aaron Lewis
Affiliation:
Division of Applied Physics, Hebrew University, Jerusalem, Israel
Leslie M. Loew
Affiliation:
Department of Physiology and Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, CT , 06030
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Extract

Confocal microscopy is an excellent high resolution method to image fluorescently labeled cells. However, the use of confocal microscopy to monitor physiological events, such as membrane potential changes, in living cells is hampered by photobleaching and phototoxicity. To reduce the collateral damage from excitation of fluorescent probes outside the optical slice, Webb and co-workers introduced the use of two-photon excited (TPE) fluorescence in laser scanning microscopy.1 Two-photon absorption depends on the square of the incident light intensity; this has the effect of confining excitation to the plane of focus where the photon flux density is greatest. The wavelength of the exciting light is in the near infrared facilitating penetration of thick tissues. Due to these significant advantages this methodology is rapidly gaining popularity as a tool for live cell and tissue imaging.

To further exploit non-linear optical processes in laser scanning microscopy, we have developed surface second harmonic generation (SHG) as a powerful new imaging modality.

Type
Advances in Multi-Photon imaging
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. 810 - 811
Copyright
Copyright © Microscopy Society of America

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References

References:

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7. This work was supported by the Office of Naval Research under Grant No. N0014-98-1-0703, National Institutes of Health GM35063, and the National Science Foundation Academic Research Infrastructure DBI-9601609.Google Scholar