Controlling cellular and organismal microenvironment using light

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The behavior of cells and organisms is intimately linked to the physical and chemical properties of their immediate microenvironment. The stiffness and adhesiveness of the substrate to which cells adhere, both in vivo and in culture, can profoundly affect cellular properties and behavior, including shape, motility, and fate choices during differentiation. Model organisms such as the nematode C. elegans use mechanosensation to integrate the topography and the texture of their environment into their behavior. The response of cells and organisms to their microenvironment can in turn be exploited to control their behaviors. For example, putting cells on linear micropatterns of extracellular matrix (ECM) can induce persistent directional migration, whereas plating them on isolated disc-shaped patterns can render them immobile. Thus, micropatterns of ECM can be used to generate stereotypical, highly reproducible cell behaviors facilitating systematic quantitative analysis of underlying mechanisms in live-cell fluorescence microscopy studies. Similarly, by placing C. elegans in defined 3D microenvironments, their behavior can be controlled and their neuronal responses to topographical features and chemical cues be determined. We propose to establish a photopatterning system in McGill's Life Sciences Complex for three main applications: (1) micropatterning of ECM to control cell adhesion and migration behavior, (2) microfabrication of complex polydimethylsiloxane (PDMS) 3D structures for microfluidics or to study how cells and C. elegans respond to complex 3D topography and (3) structuring hydrogels to mimic complex 3D tissue environments that cells encounter. To establish this photopatterning system, we are requesting a “PRIMO” UV photopatterning system (Alvéole, France), as well as components to enable its integration into an existing, currently under-utilized inverted light microscopy system. The PRIMO system will be placed under the management of the Advanced BioImaging Facility (ABIF), who will provide training to users and support the instrument, making micropatterning, microstructuring, and microfabrication highly accessible to trainees and researchers. The PRIMO system is essential for our NSERC-funded research programs in diverse research areas including single-molecule biophysics, focal adhesion dynamics/cell migration, collective cell migration/morphogenesis, immune cell dynamics, neural circuit function and computational modeling. Controlling the cellular and organismal microenvironment is an emerging key aspect of experimental design enabling new questions to be addressed with applications in basic science, organoid biology, tissue engineering, and regenerative medicine. Thus, having this system in place will significantly enhance HQP training. While six groups are included in the current application, the equipment will be accessible to >100 labs in the Life Sciences Complex and the broader research community through the ABIF.