Development of quantitative, high spatiotemporal resolution imaging methods

Over the last ten years, enormous developments in imaging technologies, quantitative analysis algorithms, new optical probes and breakthrough genetic engineering methods have enabled the rapid emergence and application of novel single molecule-based fluorescence techniques, such as single particle tracking and super-resolution nanoscopy. Yet, important challenges remain in pushing the application of these methods in biology. One of these limitations is the ability to extract quantitative information from the images, which is confounded by fluorophore photophysics. In addition, the temporal resolution is also limited by fluorophore photostability and photoswitching characteristics.

We are developing different methods to circumvent these limitations and extend the spatiotemporal resolution and quantitative power of super-resolution microscopy and single molecule imaging. In particular, we focus on different fronts: a) developing new multi-colour single molecule imaging methodologies that allow us to map real-time interactions between different molecules and their environment in living cells with nanometre precision over multiple temporal scales; b) analysis algorithms that provide detailed quantification of multicolour single particle tracking data to reveal diffusion patterns and dynamic interactions between individual molecules in living cells; c) quantitative algorithms (including deep-learning) applied to images obtained by different multi-colour super-resolution methods (STORM, DNA-PAINT & STED).