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Simultaneous spectral illumination of microplates for high-throughput optogenetics and photobiology.
Vogt, Arend; Paulat, Raik; Parthier, Daniel; Just, Verena; Szczepek, Michal; Scheerer, Patrick; Xu, Qianzhao; Möglich, Andreas; Schmitz, Dietmar; Rost, Benjamin R; Wenger, Nikolaus.
Afiliación
  • Vogt A; Department of Neurology with Experimental Neurology, Translational Neuromodulation Group, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Paulat R; Neuroscience Research Center, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Parthier D; Department of Neurology with Experimental Neurology, Translational Neuromodulation Group, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Just V; Faculty of Energy and Information, HTW-Berlin University for Applied Sciences, D-10318 Berlin, Germany.
  • Szczepek M; Neuroscience Research Center, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Scheerer P; Department of Neurology with Experimental Neurology, Translational Neuromodulation Group, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Xu Q; Faculty of Energy and Information, HTW-Berlin University for Applied Sciences, D-10318 Berlin, Germany.
  • Möglich A; Institute of Medical Physics and Biophysics, Group Structural Biology of Cellular Signaling, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Schmitz D; Institute of Medical Physics and Biophysics, Group Structural Biology of Cellular Signaling, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, D-10117 Berlin, Germany.
  • Rost BR; Department of Biochemistry, University of Bayreuth, D-95447 Bayreuth, Germany.
  • Wenger N; Department of Biochemistry, University of Bayreuth, D-95447 Bayreuth, Germany.
Biol Chem ; 2024 Sep 23.
Article en En | MEDLINE | ID: mdl-39303162
ABSTRACT
The biophysical characterization and engineering of optogenetic tools and photobiological systems has been hampered by the lack of efficient methods for spectral illumination of microplates for high-throughput analysis of action spectra. Current methods to determine action spectra only allow the sequential spectral illumination of individual wells. Here we present the open-source RainbowCap-system, which combines LEDs and optical filters in a standard 96-well microplate format for simultaneous and spectrally defined illumination. The RainbowCap provides equal photon flux for each wavelength, with the output of the LEDs narrowed by optical bandpass filters. We validated the RainbowCap for photoactivatable G protein-coupled receptors (opto-GPCRs) and enzymes for the control of intracellular downstream signaling. The simultaneous, spectrally defined illumination provides minimal interruption during time-series measurements, while resolving 10 nm differences in the action spectra of optogenetic proteins under identical experimental conditions. The RainbowCap is also suitable for studying the spectral dependence of light-regulated gene expression in bacteria, which requires illumination over several hours. In summary, the RainbowCap provides high-throughput spectral illumination of microplates, while its modular, customizable design allows easy adaptation to a wide range of optogenetic and photobiological applications.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Biol Chem Asunto de la revista: BIOQUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Alemania
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Biol Chem Asunto de la revista: BIOQUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Alemania