An In Toto Approach to Dissecting Cellular Interactions in Complex Tissues

Area of Science:

• Cell Biology
• Developmental Biology
• Systems Biology

Background:

• Single-cell measurements reveal biological heterogeneity but are often limited to observational studies.
• Current perturbation methods typically assay endpoints after biological events, lacking real-time dynamic analysis.

Purpose of the Study:

• To introduce ShootingStar, a platform enabling automated, real-time optical perturbations for in vivo single-cell analysis.
• To demonstrate ShootingStar's capability in dissecting cellular mechanisms underlying development, morphogenesis, and polarity.

Main Methods:

• ShootingStar combines live imaging, real-time image analysis, and automated optical perturbations.
• The platform quantifies sample states to identify target cells for perturbation and validate impacts.
• Applications were demonstrated in Danio rerio lateral line development and Caenorhabditis elegans embryogenesis.

Main Results:

• ShootingStar facilitates targeted, dynamic perturbations within complex biological systems.
• The platform provides quantitative data for automated cell targeting and perturbation validation.
• Successful application in model organisms highlights its utility in studying development and polarity.

Conclusions:

• ShootingStar offers a robust approach for advanced single-cell perturbation studies in vivo.
• The platform enhances the informativeness and efficiency of analyzing cellular dynamics in complex tissues.
• Future extensions can incorporate diverse optical manipulations for broader biological investigations.