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PMID: 10585956 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

High-resolution nonlinear optical imaging of live cells by second harmonic generation.

Biophysical journal ·Vol. 77 ·No. 6 ·1999-12-00 ·Pages 3341-9

Campagnola PJ, Wei MD, Lewis A, Loew LM

Abstract

By adapting a laser scanning microscope with a titanium sapphire femtosecond pulsed laser and transmission optics, we are able to produce live cell images based on the nonlinear optical phenomenon of second harmonic generation (SHG). Second harmonic imaging (SHIM) is an ideal method for probing membranes of living cells because it offers the high resolution of nonlinear optical microscopy with the potential for near-total avoidance of photobleaching and phototoxicity. The technique has been implemented on three cell lines labeled with membrane-staining dyes that have large nonlinear optical coefficients. The images can be obtained within physiologically relevant time scales. Both achiral and chiral dyes were used to compare image formation for the case of single- and double-leaflet staining, and it was found that chirality plays a significant role in the mechanism of contrast generation. It is also shown that SHIM is highly sensitive to membrane potential, with a depolarization of 25 mV resulting in an approximately twofold loss of signal intensity.

MeSH Terms
3T3 Cells Animals Biophysical Phenomena Biophysics Cells/cytology Coloring Agents Lasers Membrane Potentials Mice Microscopy/instrumentation Nonlinear Dynamics Optics and Photonics/instrumentation Tumor Cells, Cultured
Chemicals
Coloring Agents
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Campagnola P J
Department of Physiology and Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, Connecticut 06030 USA. campagno@panda.uchc.edu
Wei M D
Lewis A
Loew L M
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(99)77165-1
Email
campagno@panda.uchc.edu
Published
1999-12-00
Pages
3341-9
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1300605
Subset
IM
Grants
NIGMS NIH HHS · 5 R01-GM35063 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(99)77165-1 PMC Full Text View journal details
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