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  LED Thermo Flow - Combining Optogenetics with Flow Cytometry

Brenker, K., Osthof, K., Yang, J., & Reth, M. (2016). LED Thermo Flow - Combining Optogenetics with Flow Cytometry. Journal of visualized experiments, 118, e54707-e54707. doi:10.3791/54707.

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Datensatz-Permalink: https://hdl.handle.net/11858/00-001M-0000-002C-A8CB-8 Versions-Permalink: https://hdl.handle.net/21.11116/0000-0008-9AF7-4
Genre: Zeitschriftenartikel

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https://www.jove.com/de/t/54707/led-thermo-flow-combining-optogenetics-with-flow-cytometry (Verlagsversion)
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 Urheber:
Brenker, Kathrin1, 2, 3, Autor
Osthof, Kerstin1, 4, Autor
Yang, Jianying1, 3, Autor
Reth , Michael1, 3, 5, Autor
Affiliations:
1Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, 79108 Freiburg, DE, ou_2243640              
2Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, Germany, ou_persistent22              
3Centre for Biological Signaling Studies, BIOSS, University of Freiburg, Freiburg, Germany, ou_persistent22              
4Albert-Ludwigs-Universität, Freiburg, Germany, ou_persistent22              
5Insitute of Biology III (Mol. Immunology), Albert-Ludwigs-Universität, Freiburg, Germany, ou_persistent22              

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Schlagwörter: -
 Zusammenfassung: Optogenetic tools allow isolated, functional investigations of almost any signaling molecule within complex signaling pathways. A major obstacle is the controlled delivery of light to the cell sample and hence the most popular tools for optogenetic studies are microscopy-based cell analyses and in vitro experiments. The flow cytometer has major advantages over a microscope, including the ability to rapidly measure thousands of cells at single cell resolution. However, it is not yet widely used in optogenetics. Here, we present a device that combines the power of optogenetics and flow cytometry: the LED Thermo Flow. This device illuminates cells at specific wavelengths, light intensities and temperatures during flow cytometric measurements. It can be built at low cost and be used with most common flow cytometers. To demonstrate its utility, we characterized the photoswitching kinetics of Dronpa proteins in vivo and in real time. This protocol can be adapted to almost all optically controlled substances and substantially expands the set of possible experiments. More importantly, it will greatly simplify the discovery and development of new optogenetic tools.

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Sprache(n): eng - English
 Datum: 2016-12
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.3791/54707
 Art des Abschluß: -

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Titel: Journal of visualized experiments
  Andere : Journal of visualized experiments: JoVE
  Kurztitel : J. Vis. Exp.
Genre der Quelle: Zeitschrift
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: Rockville Pike, Bethesda MD : JoVE
Seiten: - Band / Heft: 118 Artikelnummer: - Start- / Endseite: e54707 - e54707 Identifikator: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X