Title data
Vogt, Arend ; Paulat, Raik ; Parthier, Daniel ; Just, Verena ; Szczepek, Michal ; Scheerer, Patrick ; Xu, Qianzhao ; Möglich, Andreas ; Schmitz, Dietmar ; Rost, Benjamin R. ; Wenger, Nikolaus:
Simultaneous spectral illumination of microplates for high-throughput optogenetics and photobiology.
In: Biological Chemistry.
Vol. 405
(2024)
Issue 11/12
.
- pp. 751-763.
ISSN 1437-4315
DOI: https://doi.org/10.1515/hsz-2023-0205
Abstract in another language
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.
Further data
Item Type: | Article in a journal |
---|---|
Refereed: | Yes |
Keywords: | action spectra; cyclic mononucleotides; GPCR; photoactivated nucleotidyl cyclases; rhodopsin; signal transduction |
Institutions of the University: | Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry II - Photobiochemistry > Chair Biochemistry II - Photobiochemistry - Univ.-Prof. Dr. Andreas Möglich Faculties Faculties > Faculty of Biology, Chemistry and Earth Sciences Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry II - Photobiochemistry |
Result of work at the UBT: | Yes |
DDC Subjects: | 500 Science > 570 Life sciences, biology |
Date Deposited: | 08 Oct 2024 06:35 |
Last Modified: | 24 Jan 2025 11:09 |
URI: | https://eref.uni-bayreuth.de/id/eprint/90590 |