On the Need of New Analysis Tools for the Quantification of Molecular Clustering in Super Resolution Pointillist Data Sets

November 8th, 2017 JULIETTE GRIFFIE King’s College London

Until recently, light microscopy has been limited to a resolution of around 200nm due the diffraction of light, in turn limiting the analysis of the resulting images to a coarse overview of the biological structures studied. To answer the increasing need of a precise insight into cellular machinery, a new family of fluorescence microscopy techniques, super-resolution, has arisen. Single molecule localisation microscopy (SMLM) in particular, allows the localisation of fluorophores with a precision of 10-30 nm, hence revealing the cells’ nanoscale architecture at the molecular level. Relying on such imaging techniques, it has been confirmed that modification of the molecular spatio-temporal organisation at (and proximal to) cell membranes, such as molecular clustering, is an important component of cell signalling. We will present novel model-based Bayesian cluster analysis techniques suitable for 2D SMLM as well as their extension to 3D and live imaging following the latest technical advancements in the field. We show the importance of such quantification tools to reach SMLM’s full potential in the context of cellular biology, allowing the transition from limited visual inspection of the data to the extraction of quantitative descriptors. We further discuss the importance of clustering in T cell activation and the resulting triggering of an effective immune response.

Seminar, November 8, 2017, 12:00. Seminar Room

Hosted by Prof. María García-Parajo