Wide-field three-photon excitation in biological samples

查看全文 作　　者：Christopher J [1]Rowlands;Demian [2]Park;Oliver T [3]Bruns;Kiryl D [2]Piatkevich;Dai [4]Fukumura;Rakesh K [4]Jain;Moungi G [3]Bawendi;Edward S [1,2,5]Boyden;Peter TC [1,6]So 机构地区：[1]Department of Biological Engineering,Massachusetts Institute of Technology,Cambridge,MA 02139,USA;[2]Media Lab,Massachusetts Institute of Technology,Cambridge,MA 02139,USA;[3]Department of Chemistry,Massachusetts Institute of Technology,Cambridge,MA 02139,USA;[4]Edwin L.Steele Laboratories,Department of Radiation Oncology,Massachusetts General Hospital and Harvard Medical School,Boston,MA 02114,USA;[5]Department of Brain and Cognitive Sciences,McGovern Institute and MIT Center for Neurobiological Engineering,Massachusetts Institute of Technology,Cambridge,MA 02139-4307,USA;[6]Department of Mechanical Engineering,Massachusetts Institute of Technology,Cambridge,MA 02139,USA 出　　处：《Light(Science & Applications)》索引2016年第5卷第1期,共9页 基　　金：support from NIH-5-P41-EB015871-27,DP3-DK10102401,1-U01-NS090438-01,1-R01-EY017656-0,6A1,1-R01-HL121386-01A1;the Biosym IRG of Singapore-MIT Alliance Research and Technology Center;the Koch Institute for Integrative Cancer Research Bridge Initiative;the Hamamatsu Inc.,and the Samsung GRO program;supported by the Wellcome Trust 093831/Z/10/Z;funding from NIH 1R24MH106075,NIH 2R01DA029639,NIH 1R01MH103910,NIH 1R01GM104948,the MIT Media Lab,the New York Stem Cell Foundation-Robertson Award and NSF CBET 1053233;an EMBO Longterm Fellowship to carry out this research;support from NIH 5U54 CA151884-04 and 9-P41-EB015871-26A1;for NCI grants R35 CA197743 and P01 CA080124 to carry out this work. 摘　　要：Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two-and three-photon microscopy.Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown,and a penetration depth of more than 700μm into fixed scattering brain tissue is achieved,approximately twice as deep as that achieved using two-photon wide-field excitation.Compatibility with live animal experiments is confirmed by imaging the cerebral vasculature of an anesthetized mouse;a complete focal stack was obtained without any evidence of photodamage.As an additional validation of the utility of wide-field three-photon excitation,functional excitation is demonstrated by performing threephoton optogenetic stimulation of cultured mouse hippocampal neurons expressing a channelrhodopsin;action potentials could reliably be excited without causing photodamage. 关 键 词：BIOPHOTONICS multiphoton microscopy OPTOGENETICS temporal focusing three-photon