Long-term in vivo single-cell lineage tracing of deep structures using three-photon activation

Light, Science & Applications
Isil TekeliDobryna Zalvidea

Abstract

Genetic labeling techniques allow for noninvasive lineage tracing of cells in vivo. Two-photon inducible activators provide spatial resolution for superficial cells, but labeling cells located deep within tissues is precluded by scattering of the far-red illumination required for two-photon photolysis. Three-photon illumination has been shown to overcome the limitations of two-photon microscopy for in vivo imaging of deep structures, but whether it can be used for photoactivation remains to be tested. Here we show, both theoretically and experimentally, that three-photon illumination overcomes scattering problems by combining longer wavelength excitation with high uncaging three-photon cross-section molecules. We prospectively labeled heart muscle cells in zebrafish embryos and found permanent labeling in their progeny in adult animals with negligible tissue damage. This technique allows for a noninvasive genetic manipulation in vivo with spatial, temporal and cell-type specificity, and may have wide applicability in experimental biology.

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Citations

Jan 18, 2018·Chembiochem : a European Journal of Chemical Biology·Weiting ZhangDavid Bensimon
Jul 3, 2019·Journal of Clinical Medicine·Sagar S Varankar, Sharmila A Bapat

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BETA
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