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Rapid image acquisition in multi-photon excitation fluorescence microscopy

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In conventional two-photon excitation fluorescence microscopy the excitation light is tightly focussed in the object plane in order to obtain the high field intensities needed for multi-photon excitation of fluorophores. However, a rough calculation suggested that focussing might not be necessary for many practical uses of multi-photon fluorescence microscopy. Here, we present experimental evidence that two-photon excitation widefield epifluorescence microscopy is indeed feasible using Ti-sapphire-laser excitation at λ = 760 nm. Finally, we discuss the possibilities of using a Free-Electron-Laser (FEL) as the excitation light source in two-photon epifluorescence microscopy.

Document Type: Research Article

DOI: http://dx.doi.org/10.1078/0030-4026-00339

Affiliations: Kirchhoff-Institute for Physics, Im Neuenheimer Feld 227, 69120 Heidelberg, Germany

Publication date: June 1, 2004

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