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Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish

Buckley, C.; Carvalho, M.T.; Young, L.K.; Rider, S.A.; McFadden, C.; Berlage, C.; Verdon, R.F.; Taylor, J.M.; Girkin, J.M.; Mullins, J.J.

Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish Thumbnail


Authors

C. Buckley

M.T. Carvalho

S.A. Rider

C. McFadden

C. Berlage

R.F. Verdon

J.M. Taylor

J.J. Mullins



Abstract

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.

Citation

Buckley, C., Carvalho, M., Young, L., Rider, S., McFadden, C., Berlage, C., …Mullins, J. (2017). Precise spatio-temporal control of rapid optogenetic cell ablation with mem-KillerRed in Zebrafish. Scientific Reports, 7(1), Article 5096. https://doi.org/10.1038/s41598-017-05028-2

Journal Article Type Article
Acceptance Date May 23, 2017
Online Publication Date Jul 11, 2017
Publication Date Jul 11, 2017
Deposit Date Jul 13, 2017
Publicly Available Date Jul 13, 2017
Journal Scientific Reports
Publisher Nature Research
Peer Reviewed Peer Reviewed
Volume 7
Issue 1
Article Number 5096
DOI https://doi.org/10.1038/s41598-017-05028-2

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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright 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/.





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