Buckley, C. and Carvalho, M. T. and Young, L. K. and Rider, S. A. and McFadden, C. and Berlage, C. and Verdon, R. F. and Taylor, J. M. and Girkin, J. M. and Mullins, J. J. (2017) 'Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish.', Scientific reports., 7 (1). p. 5096.
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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|Publisher Web site:||https://doi.org/10.1038/s41598-017-05028-2|
|Publisher statement:||Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.|
|Date accepted:||23 May 2017|
|Date deposited:||13 July 2017|
|Date of first online publication:||11 July 2017|
|Date first made open access:||13 July 2017|
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