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A 340/380 nm Light Emitting Diode Illuminator For Fura-2 AM Ratiometric Ca2+ Imaging Of Live Cells With Better Than 5 nM Precision

Peter W. Tinning, Aimee J. P.M. Franssen, Shehla U. Hridi, Trevor J. Bushell, Gail McConnell
doi: https://doi.org/10.1101/138495
Peter W. Tinning
1Centre for Biophotonics, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 ORE, UK
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  • For correspondence: peter.tinning@strath.ac.uk
Aimee J. P.M. Franssen
2Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 ORE, UK
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Shehla U. Hridi
2Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 ORE, UK
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Trevor J. Bushell
2Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 ORE, UK
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Gail McConnell
1Centre for Biophotonics, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 ORE, UK
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Abstract

We report the first demonstration of a fast wavelength-switchable 340/380 nm light emitting diode (LED) illuminator for Fura-2 ratiometric Ca2+ imaging of live cells. The LEDs closely match the excitation peaks of bound and free Fura-2 and enables the precise detection of cytosolic Ca2+ concentrations, which is only limited by the Ca2+ response of Fura-2. Using this illuminator, we have shown that Fura-2 acetoxymethyl ester (AM) concentrations as low as 250 nM can be used to detect induced Ca2+ events in tsA-201 cells and while utilizing the 150 µs switching speeds available, it was possible to image spontaneous Ca2+ transients in hippocampal neurons at a rate of 24.39 Hz that were blunted or absent at typical 0.5 Hz acquisition rates. Overall, the sensitivity and acquisition speeds available using this LED illuminator significantly improve the temporal resolution that can be obtained in comparison to current systems and supports optical imaging of fast Ca2+ events using Fura-2.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted May 16, 2017.
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A 340/380 nm Light Emitting Diode Illuminator For Fura-2 AM Ratiometric Ca2+ Imaging Of Live Cells With Better Than 5 nM Precision
Peter W. Tinning, Aimee J. P.M. Franssen, Shehla U. Hridi, Trevor J. Bushell, Gail McConnell
bioRxiv 138495; doi: https://doi.org/10.1101/138495
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A 340/380 nm Light Emitting Diode Illuminator For Fura-2 AM Ratiometric Ca2+ Imaging Of Live Cells With Better Than 5 nM Precision
Peter W. Tinning, Aimee J. P.M. Franssen, Shehla U. Hridi, Trevor J. Bushell, Gail McConnell
bioRxiv 138495; doi: https://doi.org/10.1101/138495

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