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10 March 2015 Bringing the light to high throughput screening: use of optogenetic tools for the development of recombinant cellular assays
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Abstract
The use of light-activated proteins represents a powerful tool to control biological processes with high spatial and temporal precision. These so called “optogenetic” technologies have been successfully validated in many recombinant systems, and have been widely applied to the study of cellular mechanisms in intact tissues or behaving animals; to do that, complex, high-intensity, often home-made instrumentations were developed to achieve the optimal power and precision of light stimulation. In our study we sought to determine if this optical modulation can be obtained also in a miniaturized format, such as a 384-well plate, using the instrumentations normally dedicated to fluorescence analysis in High Throughput Screening (HTS) activities, such as for example the FLIPR (Fluorometric Imaging Plate Reader) instrument. We successfully generated optogenetic assays for the study of different ion channel targets: the CaV1.3 calcium channel was modulated by the light-activated Channelrhodopsin-2, the HCN2 cyclic nucleotide gated (CNG) channel was modulated by the light activated bPAC adenylyl cyclase, and finally the genetically encoded voltage indicator ArcLight was efficiently used to measure potassium, sodium or chloride channel activity. Our results showed that stable, robust and miniaturized cellular assays can be developed using different optogenetic tools, and efficiently modulated by the FLIPR instrument LEDs in a 384-well format. The spatial and temporal resolution delivered by this technology might enormously advantage the early stages of drug discovery, leading to the identification of more physiological and effective drug molecules.
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Viviana Agus, Alberto Di Silvio, Jean Francois Rolland, Anna Mondini, Sara Tremolada, Katharina Montag, Lia Scarabottolo, Loredana Redaelli, and Stefan Lohmer "Bringing the light to high throughput screening: use of optogenetic tools for the development of recombinant cellular assays", Proc. SPIE 9305, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics II, 93052T (10 March 2015); https://doi.org/10.1117/12.2077579
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