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- Title
A Bright and Fast Red Fluorescent Protein Voltage Indicator That Reports Neuronal Activity in Organotypic Brain Slices.
- Authors
Abdelfattah, Ahmed S.; Farhi, Samouil L.; Yongxin Zhao; Brinks, Daan; Peng Zou; Ruangkittisakul, Araya; Platisa, Jelena; Pieribone, Vincent A.; Ballanyi, Klaus; Cohen, Adam E.; Campbell, Robert E.
- Abstract
Optical imaging of voltage indicators based on green fluorescent proteins (FPs) or archaerhodopsin has emerged as a powerful approach for detecting the activity of many individual neurons with high spatial and temporal resolution. Relative to green FP-based voltage indicators, a bright red-shifted FP-based voltage indicator has the intrinsic advantages of lower phototoxicity, lower autofluorescent background, and compatibility with blue-light-excitable channelrhodopsins. Here, we report a bright red fluorescent voltage indicator (fluorescent indicator for voltage imaging red; FlicRl) with properties that are comparable to the best available green indicators. To develop FlicRl, we used directed protein evolution and rational engineering to screen libraries of thousands of variants. FlicR 1 faithfully reports single action potentials (~ 3% AF IF ) and tracks electrically driven voltage oscillations at 100 Hz in dissociated Sprague Dawley rat hippocampal neurons in single trial recordings. Furthermore, FlicRl can be easily imaged with wide-field fluorescence microscopy. We demonstrate that FlicRl can be used in conjunction with a blue-shifted channelrhodopsin for all-optical electrophysiology, although blue light photoactivation of the FlicRl chromophore presents a challenge for applications that require spatially overlapping yellow and blue excitation.
- Subjects
GREEN fluorescent protein; RHODOPSIN; NEURONS; BRAIN; NEUROSCIENCES
- Publication
Journal of Neuroscience, 2016, Vol 36, Issue 8, p2458
- ISSN
0270-6474
- Publication type
Article
- DOI
10.1523/JNEUROSCI.3484-15.2016