Bègue Aurélien, Papagiakoumou Eirini, Leshem Ben, Conti Rossella, Enke Leona, Oron Dan, Emiliani Valentina
Wavefront-engineering Microscopy Group, Neurophysiology and New Microscopies Laboratory, Paris Descartes University, 45 rue des Saints-Pères 75270 Paris Cedex 06, France.
Department of physics of Complex Systems, Weizmann Institute of Science, Rehovot 76100, Israel.
Biomed Opt Express. 2013 Nov 18;4(12):2869-79. doi: 10.1364/BOE.4.002869. eCollection 2013.
The use of wavefront shaping to generate extended optical excitation patterns which are confined to a predetermined volume has become commonplace on various microscopy applications. For multiphoton excitation, three-dimensional confinement can be achieved by combining the technique of temporal focusing of ultra-short pulses with different approaches for lateral light shaping, including computer generated holography or generalized phase contrast. Here we present a theoretical and experimental study on the effect of scattering on the propagation of holographic beams with and without temporal focusing. Results from fixed and acute cortical slices show that temporally focused spatial patterns are extremely robust against the effects of scattering and this permits their three-dimensionally confined excitation for depths more than 500 µm. Finally we prove the efficiency of using temporally focused holographic beams in two-photon stimulation of neurons expressing the red-shifted optogenetic channel C1V1.
利用波前整形来生成局限于预定体积的扩展光学激发图案，在各种显微镜应用中已变得很常见。对于多光子激发，通过将超短脉冲的时间聚焦技术与不同的横向光整形方法（包括计算机生成全息术或广义相衬）相结合，可以实现三维限制。在此，我们展示了一项关于散射对有或没有时间聚焦的全息光束传播影响的理论和实验研究。固定和急性皮质切片的结果表明，时间聚焦的空间图案对散射效应具有极强的鲁棒性，这使得它们能够对超过500微米深度进行三维受限激发。最后，我们证明了在对表达红移光遗传学通道C1V1的神经元进行双光子刺激时使用时间聚焦全息光束的效率。
