Actin polymerisation and crosslinking drive left-right asymmetry in single cell and cell collectives.

Deviations from mirror symmetry in the development of bilateral organisms are common but the mechanisms of initial symmetry breaking are insufficiently understood. The actin cytoskeleton of individual cells self-organises in a chiral manner, but the molecular players involved remain essentially unidentified and the relationship between chirality of an individual cell and cell collectives is unclear. Here, we analysed self-organisation of the chiral actin cytoskeleton in individual cells on circular or elliptical patterns, and collective cell alignment in confined microcultures. Screening based on deep-learning analysis of actin patterns identified actin polymerisation regulators, depletion of which suppresses chirality (mDia1) or reverses chirality direction (profilin1 and CapZß). The reversed chirality  is mDia1-independent but requires the function of actin-crosslinker α-actinin1. A robust correlation between the effects of a variety of actin assembly regulators on chirality of individual cells and cell collectives is revealed. Thus, actin-driven cell chirality may underlie tissue and organ asymmetry.

An integrated platform to facilitate the calculation, validation and visualization of optical flow velocities in biological images.

Optical flow algorithms have seen poor adoption in the biological community compared with particle image velocimetry for quantifying cellular dynamics because of the lack of proper validation and an intuitive user interface. To address these challenges, we present OpFlowLab, an integrated platform that integrates our motion estimation workflow. Using routines in our workflow, we demonstrate that optical flow algorithms are more accurate than PIV in simulated images of the movement of nuclei. Qualitative assessment with actual nucleus images further supported this finding. Additionally, we show that refinement of the optical flow velocities is possible with a simple object-matching procedure, opening up the possibility of obtaining reasonable velocity estimates under less ideal imaging conditions. To visualize velocity fields, we employ artificial tracers to allow for the drawing of pathlines. Through the adoption of OpFlowLab, we are confident that optical flow algorithms will allow for the exploration of dynamic biological systems in greater accuracy and detail.