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

Ariane Deniset-Besseau (1, 2) , Sandrine Leveque-Fort (1, 2) , M.P. Fontaine-Aupart (1, 2) , Gérard Roger (3) , Patrick Georges (3)

1 PPM - Laboratoire de Photophysique Moléculaire
2 CPBM - Centre de Photonique Biomédicale
3 Laboratoire Charles Fabry de l'Institut d'Optique / Elsa

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, timegated 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

Domains: Optics [physics.optics]

Ariane Deniset-Besseau, Sandrine Leveque-Fort, M.P. Fontaine-Aupart, Gérard Roger, Patrick Georges. Three-dimensional time-resolved fluorescence imaging by multifocal multiphoton microscopy for a photosensitizer study in living cells. Applied optics, 2007, 46 (33), pp.8045-8051. ⟨hal-00691865⟩