PUBLICATION

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

Authors: Tekeli, I., Aujard, I., Trepat, X., Jullien, L., Raya, A., Zalvidea, D.
ID: ZDB-PUB-180902-2
Date: 2016
Source: Light, science & applications 5: e16084 (Journal)
Registered Authors: Raya, Angel, Tekeli, Isil
Keywords: multi-photon microscopy, photoactivation, three-photon microscopy, zebrafish
MeSH Terms: none
PubMed: 30167169 Full text @ Light Sci Appl

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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