Excitation beyond the monochromatic laser limit: Simultaneous 3-D confocal and 
multiphoton microscopy with a tapered fiber as white-light laser source.

Betz, Timo; Teipel, Jörn; Koch, Daniel; Härtig, Wolfgang; Guck, Jochen; Käs, Josef; Giessen , Harald

doi:10.1117/1.2114788

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Excitation beyond the monochromatic laser limit: Simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source.

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Guck, Jochen
Guck Division, Max Planck Institute for the Science of Light, Max Planck Society;
Guck Division, Max-Planck-Zentrum für Physik und Medizin, Max Planck Institute for the Science of Light, Max Planck Society;
Dept. of Physics, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Erlangen, Germany;

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Citation

Betz, T., Teipel, J., Koch, D., Härtig, W., Guck, J., Käs, J., et al. (2001). Excitation beyond the monochromatic laser limit: Simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source. Journal of Biomedical Optics, 10(5): 054009. doi:10.1117/1.2114788.

Cite as: https://hdl.handle.net/21.11116/0000-000F-1DBE-C

Abstract

Confocal and multiphoton microscopy are essential tools in modern life sciences. They allow fast and highly resolved imaging of a
steadily growing number of ﬂuorescent markers, ranging from ﬂuorescent proteins to quantum dots and other ﬂuorophores, used for the localization of molecules and the quantitative detection of molecular
properties within living cells and organisms. Up to now, only one physical limitation seemed to be unavoidable. Both confocal and
multiphoton microscopy rely on lasers as excitation sources, and their monochromatic radiation allows only a limited number of simultaneously usable dyes, which depends on the speciﬁc number of laser lines available in the used microscope. We have overcome this limitation by successfully replacing all excitation lasers in a standard confocal microscope with pulsed white light ranging from 430 to 1300 nm generated in a tapered silica ﬁber. With this easily reproducible method, simultaneous confocal and multiphoton microscopy was demonstrated. By developing a coherent and intense laser source with spectral width comparable to a mercury lamp, we provide the ﬂexibility to excite any desired ﬂuorophore combination