From cell mechanics to light sheet microscopy: imaging forces, molecules, cells and embryos at high spatiotemporal resolution
By definition, living specimens are animate. Therefore, a full understanding of dynamic biological systems will only be obtained by observing them with enough 4D spatio-temporal resolution and for a sufficient duration, to capture the phenomena of interest. I will first present efforts toward understanding how mechanical forces drive cell protrusion, tissue formation and matrix remodeling within 3D environments. By utilizing synthetic hydrogels and finite element models, I will describe a technique to measure the traction forces that drive cell protrusion in 3D. When acting collectively, these protrusion events also drive matrix remodeling and tissue formation. To measure this process, I will describe a microfabricated platform that permits high throughput simultaneous measurement of tissue mechanics, geometry and protein conformation. However, the remodeling events that drive cell migration and tissue formation span multiple length and time scales ranging from a few microns and seconds for individual cell protrusion events to several millimeters and days for collective tissue condensation. Unfortunately, conventional widefield or confocal microscopes are either too slow, lack the spatial resolution, or induce too much photodamage to capture these phenomena in detail...