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Dev Cell. 2017 Nov 20;43(4):530-540.e4. doi: 10.1016/j.devcel.2017.10.021.

An In Toto Approach to Dissecting Cellular Interactions in Complex Tissues.

Author information

1
Developmental Biology Program, Sloan-Kettering Institute, New York, NY 10065 USA.
2
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065 USA.
3
Developmental Biology Program, Sloan-Kettering Institute, New York, NY 10065 USA. Electronic address: baoz@mskcc.org.

Abstract

Single-cell measurements have broadened our understanding of heterogeneity in biology, yet have been limited to mostly observational studies of normal or globally perturbed systems. Typically, perturbations are utilized in an open-ended approach wherein an endpoint is assayed during or after the biological event of interest. Here we describe ShootingStar, a platform for perturbation analysis in vivo, which combines live imaging, real-time image analysis, and automated optical perturbations. ShootingStar builds a quantitative record of the state of the sample being analyzed, which is used to automate the identification of target cells for perturbation, as well as to validate the impacts of the perturbation. We used ShootingStar to dissect the cellular basis of development, morphogenesis, and polarity in the lateral line of Danio rerio and the embryo of Caenorhabditis elegans. ShootingStar can be extended to diverse optical manipulations and enables more robust and informative single-cell perturbations in complex tissues.

PMID:
29161596
PMCID:
PMC5728429
DOI:
10.1016/j.devcel.2017.10.021
[Indexed for MEDLINE]
Free PMC Article

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