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

Lookup NU author(s): Professor Christopher Buckley, Dr Laura Young, Dr Jonathan Taylor

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

© 2017 The Author(s).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.


Publication metadata

Author(s): Buckley C, Carvalho MT, Young LK, Rider SA, McFadden C, Berlage C, Verdon RF, Taylor JM, Girkin JM, Mullins JJ

Publication type: Article

Publication status: Published

Journal: Scientific Reports

Year: 2017

Volume: 7

Print publication date: 01/12/2017

Online publication date: 11/07/2017

Acceptance date: 23/05/2017

Date deposited: 21/05/2020

ISSN (electronic): 2045-2322

Publisher: Nature Publishing Group

URL: https://doi.org/10.1038/s41598-017-05028-2

DOI: 10.1038/s41598-017-05028-2

PubMed id: 28698677


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