Single molecule fluorescence detection of BODIPY-FL molecules for monitoring protein synthesis |
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Authors: | K Perronet N Westbrook N Soler S Yoshizawa |
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Institution: | a Laboratoire Charles Fabry de l’Institut d’Optique, UMR CNRS 8501 et Université Paris-Sud, Institut d’Optique Graduate School, Campus Polytechnique, RD 128, 91127 Palaiseau Cedex, France b Laboratoire de Chimie et Biologie Structurales, ICSN-CNRS 1 avenue de la terrasse, 91190 Gif-sur-Yvette, France |
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Abstract: | We have observed single DNA strands labeled with a small fluorescent molecule, BODIPY-FL, using objective based total internal fluorescence microscopy. In spite of its photobleaching, this small fluorophore is potentially useful for monitoring biochemical processes like protein synthesis because it can be efficiently incorporated into proteins by the ribosome through a charged epsilon-labeled fluorescent lysine tRNA. Using evanescent wave laser excitation at 488 nm, we have been able to observe single Bodipy molecules using integration times as low as 20 ms with a good signal to noise ratio, and for several images before photobleaching. We have measured the fluorescence decay due to photobleaching and have been able to lengthen it by a factor of 5 using oxygen scavenger systems. |
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Keywords: | 87 64&minus t 42 62 |
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