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Issue 2, 2015
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Upgrading a microplate reader for photobiology and all-optical experiments

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Abstract

Automation can vastly reduce the cost of experimental labor and thus facilitate high experimental throughput, but little off-the-shelf hardware for the automation of illumination experiments is commercially available. Here, we use inexpensive open-source electronics to add programmable illumination capabilities to a multimode microplate reader. We deploy this setup to characterize light-triggered phenomena in three different sensory photoreceptors. First, we study the photoactivation of Arabidopsis thaliana phytochrome B by light of different wavelengths. Second, we investigate the dark-state recovery kinetics of the Synechocystis sp. blue-light sensor Slr1694 at multiple temperatures and imidazole concentrations; while the kinetics of the W91F mutant of Slr1694 are strongly accelerated by imidazole, the wild-type protein is hardly affected. Third, we determine the light response of the Beggiatoa sp. photoactivatable adenylate cyclase bPAC in Chinese hamster ovary cells. bPAC is activated by blue light in dose-dependent manner with a half-maximal intensity of 0.58 mW cm−2; intracellular cAMP spikes generated upon bPAC activation decay with a half time of about 5 minutes after light switch-off. Taken together, we present a setup which is easily assembled and which thus offers a facile approach to conducting illumination experiments at high throughput, reproducibility and fidelity.

Graphical abstract: Upgrading a microplate reader for photobiology and all-optical experiments

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Publication details

The article was received on 29 Sep 2014, accepted on 24 Oct 2014 and first published on 24 Oct 2014


Article type: Paper
DOI: 10.1039/C4PP00361F
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Citation: Photochem. Photobiol. Sci., 2015,14, 270-279
  • Open access: Creative Commons BY license
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    Upgrading a microplate reader for photobiology and all-optical experiments

    F. Richter, U. S. Scheib, J. Mehlhorn, R. Schubert, J. Wietek, O. Gernetzki, P. Hegemann, T. Mathes and A. Möglich, Photochem. Photobiol. Sci., 2015, 14, 270
    DOI: 10.1039/C4PP00361F

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      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
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      [Original citation] - Published by The Royal Society of Chemistry.

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