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Proceedings Paper

Molecular probes for two-photon excited fluorescence and second harmonic generation imaging of biological membranes
Author(s): Laurent Porres; Olivier Mongin; Bharath K. G. Bhatthula; Mireille H. Blanchard-Desce; Lionel Ventelon; Laurent Moreaux; T. Pons; Jerome Mertz
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Paper Abstract

Novel microscopies based on nonlinear optical (NLO) phenomena are attracting increasing interest in the biology community owing to their potentialities in the area of real-time, non-damaging imaging of biological systems. In particular, second-harmonic generation (SHG) and two-photon excited fluorescence (TPEF) are NLO phenomena that scale with excitation intensity squared, and thus give rise to an intrinsic 3-dimensional resolution when used in microscopic imaging. In this perspective, we have implemented a molecular engineering approach toward NLO-probes specifically designed for SHG and/or TPEF imaging of cellular membranes. We have designed nanoscale rod-like fluorophores showing very large TPEF cross-sections in the visible red, outperforming standard fluorophores such as fluorescein by up to two orders of magnitude. Bolaamphiphilic derivatives combining high TPEF cross-sections and affinity for cellular membranes were prepared. Their incorporation into model or cell membranes can be monitored by TPEF microscopy. Amphiphilic push-pull chromophores showing both high TPA and SHG cross-sections in the near-IR region were designed as NLO-probes for imaging of biological membranes by simultaneous SHG and TPEF microscopy. These NLO-phores offer intriguing potentialities for imaging of fundamental biological processes such as adhesion, fusion or for reporting of membrane electrical potentials.

Paper Details

Date Published: 1 November 2002
PDF: 10 pages
Proc. SPIE 4812, Nonlinear Spectroscopy, (1 November 2002); doi: 10.1117/12.451378
Show Author Affiliations
Laurent Porres, Univ. de Rennes I (France)
Olivier Mongin, Univ. de Rennes I (France)
Bharath K. G. Bhatthula, Univ. de Rennes I (France)
Mireille H. Blanchard-Desce, Univ. de Rennes I (France)
Lionel Ventelon, Ecole Normale Superieure (France)
Laurent Moreaux, Ecole Superieure de Physique et de Chimie Industrielles (France)
T. Pons, Ecole Superieure de Physique et de Chimie Industrielles (France)
Jerome Mertz, Ecole Superieure de Physique et de Chimie Industrielles (France)

Published in SPIE Proceedings Vol. 4812:
Nonlinear Spectroscopy
David L. Andrews, Editor(s)

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