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Synchrotron UV Fluorescence Microscopy Uncovers New Probes in Cells and Tissues

Published online by Cambridge University Press:  25 August 2010

Frederic Jamme
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France Cepia, Institut National de la Recherche Agronomique (INRA), BP 71627, 44316 Nantes, France
Sandrine Villette
Affiliation:
Centre de Biophysique Moléculaire, CNRS UPR4301, Rue Charles Sadron, 45071 Orléans, France
Alexandre Giuliani
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France Cepia, Institut National de la Recherche Agronomique (INRA), BP 71627, 44316 Nantes, France
Valerie Rouam
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France
Frank Wien
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France
Bruno Lagarde
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France
Matthieu Réfrégiers
Affiliation:
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France
Corresponding

Abstract

Use of deep ultraviolet (DUV, below 350 nm) fluorescence opens up new possibilities in biology because it does not need external specific probes or labeling but instead allows use of the intrinsic fluorescence that exists for many biomolecules when excited in this wavelength range. Indeed, observation of label free biomolecules or active drugs ensures that the label will not modify the biolocalization or any of its properties. In the past, it has not been easy to accomplish DUV fluorescence imaging due to limited sources and to microscope optics. Two worlds were coexisting: the spectrofluorometric measurements with full spectrum information with DUV excitation, which lacked high-resolution localization, and the microscopic world with very good spatial resolution but poor spectral resolution for which the wavelength range was limited to 350 nm. To combine the advantages of both worlds, we have developed a DUV fluorescence microscope for cell biology coupled to a synchrotron beamline, providing fine tunable excitation from 180 to 600 nm and full spectrum acquired on each point of the image, to study DUV excited fluorescence emitted from nanovolumes directly inside live cells or tissue biopsies.

Type
Biological Applications
Copyright
Copyright © Microscopy Society of America 2010

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