Two-photon single-cell optogenetic control of neuronal.pdfVIP

Two-photon single-cell optogenetic control of neuronal activity by sculpted light Bertalan K. Andrasfalvy, Boris V. Zemelman, Jianyong Tang, and Alipasha Vaziri1 Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147 Communicated by Charles V. Shank, Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, May 14, 2010 (received for review April 6, 2010) Recent advances in optogenetic techniques have generated new Results tools for controlling neuronal activity, with a wide range of neu- ChR2 Activation by Two-Photon Temporal Focusing. We took a dif- roscience applications. The most commonly used approach has ferent approach for addressing the shortcomings of both the one- been the optical activation of the light-gated ion channel channelr- photon and the two-photon method in a fundamentally unique hodopsin-2 (ChR2). However, targeted single-cell-level optogenetic way and have demonstrated reliable single neuron–specific acti- activation with temporal precessions comparable to the spike vation with temporal resolution (≤1 ms) in hippocampal slices. timing remained challenging. Here we report fast (≤1 ms), selec- This was done by effectively decoupling the lateral (i.e., the waist tive, and targeted control of neuronal activity with single-cell size) and the axial (i.e., the Rayleigh length) beam parameters for resolution in hippocampal slices. Using temporally focused laser two-photon absorption, which are usually coupled for a Gaussian pulses (TEFO) for which the axial beam profile can be controlled beam, and thereby sculpting the spatial light distribution at the independently of its lateral distribution, large numbers of channels focal plane. For a pulsed laser source, the decoupling can eff