Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging

نویسندگان

  • Shigenori Inagaki
  • Hidekazu Tsutsui
  • Kazushi Suzuki
  • Masakazu Agetsuma
  • Yoshiyuki Arai
  • Yuka Jinno
  • Guirong Bai
  • Matthew J. Daniels
  • Yasushi Okamura
  • Tomoki Matsuda
  • Takeharu Nagai
چکیده

We report development of the first genetically encoded bioluminescent indicator for membrane voltage called LOTUS-V. Since it is bioluminescent, imaging LOTUS-V does not require external light illumination. This allows bidirectional optogenetic control of cellular activity triggered by Channelrhodopsin2 and Halorhodopsin during voltage imaging. The other advantage of LOTUS-V is the robustness of a signal-to-background ratio (SBR) wherever it expressed, even in the specimens where autofluorescence from environment severely interferes fluorescence imaging. Through imaging of moving cardiomyocyte aggregates, we demonstrated the advantages of LOTUS-V in long-term imaging are attributable to the absence of phototoxicity, and photobleaching in bioluminescent imaging, combined with the ratiometric aspect of LOTUS-V design. Collectively LOTUS-V extends the scope of excitable cell control and simultaneous voltage phenotyping, which should enable applications in bioscience, medicine and pharmacology previously not possible.

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عنوان ژورنال:

دوره 7  شماره 

صفحات  -

تاریخ انتشار 2017