Literature DB >> 33522514

Building a Simple and Versatile Illumination System for Optogenetic Experiments.

Phillip Kyriakakis1, Lourdes Fernandez de Cossio2, Patrick Wade Howard3, Sivleng Kouv3, Marianne Catanho3, Vincent J Hu4, Robert Kyriakakis3, Molly E Allen3, Yunhan Ma5, Marcelo Aguilar-Rivera3, Todd P Coleman3.   

Abstract

Controlling biological processes using light has increased the accuracy and speed with which researchers can manipulate many biological processes. Optical control allows for an unprecedented ability to dissect function and holds the potential for enabling novel genetic therapies. However, optogenetic experiments require adequate light sources with spatial, temporal, or intensity control, often a bottleneck for researchers. Here we detail how to build a low-cost and versatile LED illumination system that is easily customizable for different available optogenetic tools. This system is configurable for manual or computer control with adjustable LED intensity. We provide an illustrated step-by-step guide for building the circuit, making it computer-controlled, and constructing the LEDs. To facilitate the assembly of this device, we also discuss some basic soldering techniques and explain the circuitry used to control the LEDs. Using our open-source user interface, users can automate precise timing and pulsing of light on a personal computer (PC) or an inexpensive tablet. This automation makes the system useful for experiments that use LEDs to control genes, signaling pathways, and other cellular activities that span large time scales. For this protocol, no prior expertise in electronics is required to build all the parts needed or to use the illumination system to perform optogenetic experiments.

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Year:  2021        PMID: 33522514      PMCID: PMC9119341          DOI: 10.3791/61914

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.424


  32 in total

1.  Bi-stable neural state switches.

Authors:  André Berndt; Ofer Yizhar; Lisa A Gunaydin; Peter Hegemann; Karl Deisseroth
Journal:  Nat Neurosci       Date:  2008-12-08       Impact factor: 24.884

2.  Temporally precise labeling and control of neuromodulatory circuits in the mammalian brain.

Authors:  Dongmin Lee; Meaghan Creed; Kanghoon Jung; Thomas Stefanelli; Daniel J Wendler; Won Chan Oh; Neymi Layne Mignocchi; Christian Lüscher; Hyung-Bae Kwon
Journal:  Nat Methods       Date:  2017-04-03       Impact factor: 28.547

3.  Ultraviolet-B-mediated induction of protein-protein interactions in mammalian cells.

Authors:  Remco P Crefcoeur; Ruohe Yin; Roman Ulm; Thanos D Halazonetis
Journal:  Nat Commun       Date:  2013       Impact factor: 14.919

Review 4.  Applications of Optobiology in Intact Cells and Multicellular Organisms.

Authors:  John S Khamo; Vishnu V Krishnamurthy; Savanna R Sharum; Payel Mondal; Kai Zhang
Journal:  J Mol Biol       Date:  2017-09-04       Impact factor: 5.469

5.  Cryptochrome, phytochrome, and anthocyanin production.

Authors:  A L Mancinelli; F Rossi; A Moroni
Journal:  Plant Physiol       Date:  1991-08       Impact factor: 8.340

6.  A genetically encoded photoactivatable Rac controls the motility of living cells.

Authors:  Yi I Wu; Daniel Frey; Oana I Lungu; Angelika Jaehrig; Ilme Schlichting; Brian Kuhlman; Klaus M Hahn
Journal:  Nature       Date:  2009-08-19       Impact factor: 49.962

7.  Spatiotemporal control of cell signalling using a light-switchable protein interaction.

Authors:  Anselm Levskaya; Orion D Weiner; Wendell A Lim; Christopher A Voigt
Journal:  Nature       Date:  2009-09-13       Impact factor: 49.962

8.  A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells.

Authors:  Konrad Müller; Raphael Engesser; Stéphanie Metzger; Simon Schulz; Michael M Kämpf; Moritz Busacker; Thorsten Steinberg; Pascal Tomakidi; Martin Ehrbar; Ferenc Nagy; Jens Timmer; Matias D Zubriggen; Wilfried Weber
Journal:  Nucleic Acids Res       Date:  2013-01-25       Impact factor: 16.971

9.  Optimized second-generation CRY2-CIB dimerizers and photoactivatable Cre recombinase.

Authors:  Amir Taslimi; Brian Zoltowski; Jose G Miranda; Gopal P Pathak; Robert M Hughes; Chandra L Tucker
Journal:  Nat Chem Biol       Date:  2016-04-11       Impact factor: 15.040

Review 10.  Light-Controlled Mammalian Cells and Their Therapeutic Applications in Synthetic Biology.

Authors:  Maysam Mansouri; Tobias Strittmatter; Martin Fussenegger
Journal:  Adv Sci (Weinh)       Date:  2018-09-30       Impact factor: 16.806

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