Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish.
Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish.
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DOI：
10.1038/s41598-017-05028-2
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发表时间：
2017-07-11
影响因子：
4.6
通讯作者：
Mullins JJ
中科院分区：
文献类型：
--
作者：
Buckley C;Carvalho MT;Young LK;Rider SA;McFadden C;Berlage C;Verdon RF;Taylor JM;Girkin JM;Mullins JJ
在体内杀死单个或群细胞的能力对于研究细胞过程及其生理功能是重要的。细胞特异性基因编码的光敏蛋白，如KillerRed，允许用低功率激光进行时空光遗传消融。我们报告通过使用选择性平面照明显微镜（SPIM），显著提高了斑马鱼肾脏细胞靶向的分辨率和速度。此外，通过将贝塞尔光束整合到SPIM成像臂中，我们能够提高瞄准速度和精度。贝塞尔光束的低衍射加上通过高NA透镜紧密聚焦的能力，使得在活的、发育中的斑马鱼肾脏的解剖深度上精确、快速地靶向细胞亚群。我们证明了这些特定的靶向策略显著提高了光消融的速度以及鱼的存活率。
The ability to kill individual or groups of cells in vivo is important for studying cellular processes and their physiological function. Cell-specific genetically encoded photosensitizing proteins, such as KillerRed, permit spatiotemporal optogenetic ablation with low-power laser light. We report dramatically improved resolution and speed of cell targeting in the zebrafish kidney through the use of a selective plane illumination microscope (SPIM). Furthermore, through the novel incorporation of a Bessel beam into the SPIM imaging arm, we were able to improve on targeting speed and precision. The low diffraction of the Bessel beam coupled with the ability to tightly focus it through a high NA lens allowed precise, rapid targeting of subsets of cells at anatomical depth in live, developing zebrafish kidneys. We demonstrate that these specific targeting strategies significantly increase the speed of optoablation as well as fish survival.
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影响因子：
3.1
作者：
Valentine, Ronan M.;Ibbotson, Sally H.;Moseley, Harry
通讯作者：
Moseley, Harry
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
3.5
作者：
Carpentier, Philippe;Violot, Sebastien;Bourgeois, Dominique
通讯作者：
Bourgeois, Dominique
DOI：
10.1152/ajprenal.00247.2015
发表时间：
2015-09-15
影响因子：
4.2
作者：
Rider, Sebastien A.;Mullins, Linda J.;Mullins, John J.
通讯作者：
Mullins, John J.
影响因子：
2.5
作者：
Curado, Silvia;Anderson, Ryan M.;Stainier, Didier Y. R.
通讯作者：
Stainier, Didier Y. R.