Elucidating the functional organization and dynamics of the nucleus in live cells is a major challenge in cell biology to understand gene regulation. Single-molecule based approaches have revealed themselves instrumental to this end. However, these techniques are usually hindered by the lack of optical sectioning and a limited axial range, severely limiting the density and volume of data one can acquire on single nuclei at the molecular level.
In this project, we propose to use adaptive optics and point-spread function (PSF) shaping techniques to obtain fast (millisecond-range), full volumetric, in-focus imaging of entire nuclei. With the help of a deformable mirror, we will develop oblique-plane imaging compatible with angled illumination of the sample, effectively obtaining a single-plane illumination microscopy configuration compatible with high NA objectives, and also capable of fast refocusing.
The expertise and compatibility of the partners involved assure the feasibility of this ambitious project.