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Applied Optics

Applied Optics


  • Editor: James C. Wyant
  • Vol. 46, Iss. 33 — Nov. 20, 2007
  • pp: 8045–8051

Three-dimensional time-resolved fluorescence imaging by multifocal multiphoton microscopy for a photosensitizer study in living cells

A. Deniset-Besseau, S. Lévêque-Fort, M. P. Fontaine-Aupart, G. Roger, and P. Georges  »View Author Affiliations

Applied Optics, Vol. 46, Issue 33, pp. 8045-8051 (2007)

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Two-photon fluorescence microscopy is widely applied to biology and medicine to study both the structure and dynamic processes in living cells. The main issue is the slow acquisition rate due to the point scanning approach limiting the multimodal detection ( x , y , z , t ) . To extend the performances of this powerful technique, we present a time-resolved multifocal multiphoton microscope (MMM) based on laser amplitude splitting. An array of 8 × 8 foci is created on the sample that gives a direct insight of the fluorescence localization. Four-dimensional (4D) imaging is obtained by combining simultaneous foci scanning, time-gated detection, and z displacement. We illustrate time-resolved MMM capabilities for 4D imaging of a photosensitizer inside living colon cancer cells. The aim of this study is to have a better understanding of the photophysical processes implied in the photosensitizer reactivity.

© 2007 Optical Society of America

OCIS Codes
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: July 9, 2007
Revised Manuscript: September 25, 2007
Manuscript Accepted: September 27, 2007
Published: November 19, 2007

Virtual Issues
Vol. 2, Iss. 12 Virtual Journal for Biomedical Optics

A. Deniset-Besseau, S. Lévêque-Fort, M. P. Fontaine-Aupart, G. Roger, and P. Georges, "Three-dimensional time-resolved fluorescence imaging by multifocal multiphoton microscopy for a photosensitizer study in living cells," Appl. Opt. 46, 8045-8051 (2007)

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