TY - JOUR
T1 - Multiplexed temporally focused light shaping for high-resolution multi-cell targeting
AU - Accanto, Nicolò
AU - Molinier, Clément
AU - Tanese, Dimitrii
AU - Ronzitti, Emiliano
AU - Newman, Zachary L.
AU - Wyart, Claire
AU - Isacoff, Ehud
AU - Papagiakoumou, Eirini
AU - Emiliani, Valentina
N1 - Publisher Copyright:
© 2018 Optical Society of America.
PY - 2018/11/20
Y1 - 2018/11/20
N2 - Optical wavefront shaping is a powerful technique to control the distribution of light in the focus of a microscope. This ability, combined with optogenetics, holds great promise for precise manipulation of neuronal activity with light. However, a deeper understanding of complex brain circuits requires pushing light-shaping methods into a new regime: the simultaneous excitation of several tens of targets, arbitrarily distributed in the three dimensions, with single-cell resolution. To this end, we developed a new optical scheme, based on the spatio-temporal shaping of a pulsed laser beam, to project several tens of spatially confined two-photon excitation patterns in a large volume. Compatibility with several different phase-shaping strategies allows the system to be optimized towards flexibility, simplicity, or multiple independent light manipulations, thus providing new routes for precise three-dimensional optogenetics. To validate the method, we performed multi-cell volumetric excitation of photoactivatable GCaMP in the central nervous system of drosophila larvae, a challenging structure with densely arrayed neurons, and photoconversion of the fluorescent protein Kaede in zebrafish larvae.
AB - Optical wavefront shaping is a powerful technique to control the distribution of light in the focus of a microscope. This ability, combined with optogenetics, holds great promise for precise manipulation of neuronal activity with light. However, a deeper understanding of complex brain circuits requires pushing light-shaping methods into a new regime: the simultaneous excitation of several tens of targets, arbitrarily distributed in the three dimensions, with single-cell resolution. To this end, we developed a new optical scheme, based on the spatio-temporal shaping of a pulsed laser beam, to project several tens of spatially confined two-photon excitation patterns in a large volume. Compatibility with several different phase-shaping strategies allows the system to be optimized towards flexibility, simplicity, or multiple independent light manipulations, thus providing new routes for precise three-dimensional optogenetics. To validate the method, we performed multi-cell volumetric excitation of photoactivatable GCaMP in the central nervous system of drosophila larvae, a challenging structure with densely arrayed neurons, and photoconversion of the fluorescent protein Kaede in zebrafish larvae.
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U2 - 10.1364/OPTICA.5.001478
DO - 10.1364/OPTICA.5.001478
M3 - Article
AN - SCOPUS:85058950606
SN - 2334-2536
VL - 5
SP - 1478
EP - 1491
JO - Optica
JF - Optica
IS - 11
ER -