Dynamic, volumetric super-resolution imaging of cellular organization and molecular orientation with reversible cryo-arrest technology

Molecular organization plays a crucial role in orchestrating cellular functions. However, observing this organization at single molecule level in crowded environment such as cell nucleus and at specific time points remains a big challenge.

This project aims to develop a unique super-resolution microscopy by combining selective plane excitation and volumetric imaging using multifocus microscopy, for dense single molecule imaging. Moreover, single molecule 3D orientation will be accessed by polarization analysis and will result in a better discrimination of local organization and neighboring molecules. Reversible cryo-arrest technology will be implemented to freeze molecular motion in the cell and to gain photo physical stability at low temperature.

We will use this technology to evaluate the in vivo structure and dynamics of DNA, nucleosomes and RNA Polymerase at transcriptionally active genes with unprecedented resolution, in the same cell and at consecutive time points.