New sensors and strategies for the visualization of nanoscale activity and 3D cell structure

Living systems use nanoscale organization to compartmentalize and guide the chemical processes occurring inside them. This spatiotemporal heterogeneity is thought to be a key concept for e.g. signaling specificity, yet is difficult to study as it can only be measured in the intact cell. However, at the current state of the art it is difficult to measure chemical (signaling) processes at high spatiotemporal resolution, or to measure whole-cell ultrastructuring in reasonable time scales. This project proposes to develop new optical biosensors for the visualization of nanoscale signaling heterogeneity. These probes will be compatible with super-resolution imaging by exploiting new architectures and screening methodologies. In parallel we will optimize the performance of the imaging methodology of choice, SOFI, and extend it to multi-parameter acquisitions based on pointspread- function engineering. The resulting probes and methodology will be applied to visualizing the nanoscale heterogeneity of cAMP/PKA-mediated signaling together with the surrounding context. The potential and impact of these tools will be further highlighted by collaborative applications to mitochondrial (dys-) function and neuronal plasticity, emphasizing the generic character of this work while also expanding its impact to multiple research fields.