opt sliderMultiplexed temporally focused light shaping for high-resolution multi-cell targeting

A picture from Valentina Emiliani's article, written together with her Team, has been chosen for the cover of OPTICA (Vol. 5, Issue 11, pp. 1478-1491 (2018). Credits for the cover picture to Clément Molinier.



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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