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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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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-002C-A8CB-8 Version Permalink: https://hdl.handle.net/21.11116/0000-0008-9AF7-4
Genre: Journal Article

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https://www.jove.com/de/t/54707/led-thermo-flow-combining-optogenetics-with-flow-cytometry (Publisher version)
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 Creators:
Brenker, Kathrin1, 2, 3, Author
Osthof, Kerstin1, 4, Author
Yang, Jianying1, 3, Author
Reth , Michael1, 3, 5, Author
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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 Abstract: 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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Language(s): eng - English
 Dates: 2016-12
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.3791/54707
 Degree: -

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Title: Journal of visualized experiments
  Other : Journal of visualized experiments: JoVE
  Abbreviation : J. Vis. Exp.
Source Genre: Journal
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Publ. Info: Rockville Pike, Bethesda MD : JoVE
Pages: - Volume / Issue: 118 Sequence Number: - Start / End Page: e54707 - e54707 Identifier: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X