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A NOVEL RHODOPSIN-BASED VOLTAGE INDICATOR FOR SIMULTANEOUS TWO PHOTON OPTICAL RECORDING WITH GCAMP IN VIVO
Vincent Villetteand 10 co-authors
Institut de Biologie de l’École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University
FENS Forum 2026 (2026)
Barcelona, Spain
Board PS07-10AM-067
Presentation
Date TBA
Board: PS07-10AM-067
Event Information
Poster Board
PS07-10AM-067
Poster
View posterAbstract
Genetically encoded voltage indicators (GEVIs) allow optical recording of membrane potential from targeted cells in vivo. Red-reporting GEVIs that are compatible with two photon microscopy and that can be multiplexed in vivo with green reporters like GCaMP, are currently lacking. To address this, we explored diverse rhodopsin proteins as GEVIs and engineered a novel GEVI, 2Photron-ST, based on a rhodopsin from the green algae Klebsormidium nitens. Using two photon ultrafast local volume excitation (ULoVE), 2Photron was compared, in several cell types, with a state-of-the-art green 2P GEVIs, i.e. JEDI2P-Kv, and a red 1P GEVI Voltron2-ST. We found 2Photron-ST enabled multiplexed recording of spiking and subthreshold voltages simultaneously with GCaMP6/8f calcium signals in visual cortical neurons of awake, behaving mice. These recordings revealed the cell-specific relationship of spiking and subthreshold voltage dynamics with GCaMP responses, highlighting the challenges of extracting underlying spike trains from calcium imaging.
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