Synchrotron UV fluorescence microscopy uncovers new probes in cells and tissues.
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.
Jamme
Frederic
F
Synchrotron SOLEIL, L'Orme des Merisiers, 91192 Gif sur Yvette, France.
Villette
Sandrine
S
Giuliani
Alexandre
A
Rouam
Valerie
V
Wien
Frank
F
Lagarde
Bruno
B
Réfrégiers
Matthieu
M
D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008027:Light; D008856:Microscopy, Fluorescence / Q000379:methods*; D017356:Synchrotrons; D014466:Ultraviolet Rays