Clear imaging of Förster resonance energy transfer (FRET) signals of Ras activation by a time-lapse three-dimensional deconvolution system

Authors

  • TAKESHI YOKONO,

    1. M & S Instruments Inc., 113, Yarai-cho, Shinjuku-ku, Tokyo 162-0805, Japan
      *Graduate School of Agriculture & Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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  • HIROTO KOTANIGUCHI,

    1. M & S Instruments Inc., 113, Yarai-cho, Shinjuku-ku, Tokyo 162-0805, Japan
      *Graduate School of Agriculture & Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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  • and * YASUHISA FUKUI

    1. M & S Instruments Inc., 113, Yarai-cho, Shinjuku-ku, Tokyo 162-0805, Japan
      *Graduate School of Agriculture & Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan
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Yasuhisa Fukui. Tel: +81 3 5841 5110; fax: +81 3 5841 8024; e-mail: ayfukui@mail.ecc.u-tokyo.ac.jp

Summary

A procedure for a time series of three-dimensional Förster resonance energy transfer observation of a cell has been established. It employs quantitative deconvolution and three-dimensional views of intensity-modulated displays. The development was done with Raichu, a synthetic Ras protein capable of producing Förster resonance energy transfer upon its activation, for which two-dimensional imaging has been established. This method gave much clearer images of Förster resonance energy transfer than does the usual method without deconvolution, even though the signals were relatively weak. The results suggest that this procedure is compatible with weak fluorescent light, which is prone to photobleaching.

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