1. Linearly polarized excitation enhances signals from fluorescent voltage indicators
- Author
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Simon Kheifets, Daan Brinks, William Bloxham, Adam E. Cohen, and Molecular Genetics
- Subjects
0303 health sciences ,Brightness ,Materials science ,business.industry ,Linear polarization ,Biophysics ,Action Potentials ,Articles ,Signal ,Fluorescence ,Noise (electronics) ,03 medical and health sciences ,0302 clinical medicine ,Optoelectronics ,Animals ,Indicators and Reagents ,business ,Polarization (electrochemistry) ,Coloring Agents ,030217 neurology & neurosurgery ,Excitation ,030304 developmental biology ,Voltage ,Fluorescent Dyes - Abstract
Voltage imaging in cells requires high-speed recording of small fluorescent signals, often leading to low signal/noise ratios. Because voltage indicators are membrane bound, their orientations are partially constrained by the plane of the membrane. We explored whether tuning the linear polarization of excitation light could enhance voltage indicator fluorescence. We tested a panel of dye- and protein-based voltage indicators in mammalian cells. The dye BeRST1 showed a 73% increase in brightness between the least and most favorable polarizations. The protein-based reporter ASAP1 showed a 22% increase in brightness, and QuasAr3 showed a 14% increase in brightness. In very thin neurites expressing QuasAr3, improvements were anomalously large, with a 170% increase in brightness between polarization parallel versus perpendicular to the dendrite. Signal/noise ratios of optically recorded action potentials were increased by up to 50% in neurites expressing QuasAr3. These results demonstrate that polarization control can be a facile means to enhance signals from fluorescent voltage indicators, particularly in thin neurites or in high-background environments.
- Published
- 2021
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