SUPER-RESOLVED IMAGING OF EARLY-STAGE DYNAMICS IN THE IMMUNE RESPONSE

The use of photo-activated fluorescent molecules to create long sequences of low-density, diffraction-limited images gives us the ability to achieve highly-precise molecule localizations. However, this methodology requires lengthy imaging times, resulting in poor temporal resolution. This is particularly problematic when dynamic interactions of live cells on short time scales are of interest. We consider the problem of shortening dramatically the acquisition times in superresolution microscopy down to seconds, in order to image the cellular dynamics during T-cell activation. For this purpose, we apply our newly developed method for super-resolution microscopy, SPARCOM, to real-time imaging of the immune response initiation. Despite the small number of frames used for reconstruction, we are able to achieve high-quality imaging that shows how T-cell receptors re-organize during the activation events, with respect to the cell’s surface. Thus, our work demonstrates the use of SPARCOM for imaging dynamic cellular processes well below the diffraction limit.