Two-Photon Holographic Stimulation of ReaChR.
Optogenetics provides a unique approach to remotely manipulate brain activity with light. Reaching the degree of spatiotemporal control necessary to dissect the role of individual cells in neuronal networks, some of which reside deep in the brain, requires joint progress in opsin engineering and light sculpting methods. Here we investigate for the first time two-photon stimulation of the red-shifted opsin ReaChR. We use two-photon (2P) holographic illumination to control the activation of individually chosen neurons expressing ReaChR in acute brain slices. We demonstrated reliable action potential generation in ReaChR-expressing neurons and studied holographic 2P-evoked spiking performances depending on illumination power and pulse width using an amplified laser and a standard femtosecond Ti:Sapphire oscillator laser. These findings provide detailed knowledge of ReaChR's behavior under 2P illumination paving the way for achieving in depth remote control of multiple cells with high spatiotemporal resolution deep within scattering tissue.
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Emmanuelle
E
Chaigneau
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Emiliano
E
Ronzitti
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Marta A
MA
Gajowa
Allen Institute for Brain Science Seattle, WA, USA.
Gilberto J
GJ
Soler-Llavina
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Dimitrii
D
Tanese
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Anthony Y B
AY
Brureau
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes UniversityParis, France; Institut National de la Santé et de la Recherche Médicale (INSERM)Paris, France.
Eirini
E
Papagiakoumou
Allen Institute for Brain Science Seattle, WA, USA.
Hongkui
H
Zeng
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, Centre National de la Recherche Scientifique UMR8250, Paris Descartes University Paris, France.
Valentina
V
Emiliani
2-photon excitation; action-potential generation; computer generated holography; cortex; neuroscience; opsin; optogenetics