Protocol for producing 3D super-resolution images

Protocol for producing 3D super-resolution images of whole chromosomes The focus here is to bring imaging of metaphase chromosomes to a new level of resolution and control. The condensed chromosomes will be flown in our microfluidic chamber (WP2) and captured out of solution using the specific chromosome-microsphere handles developed in task 3.1. We will apply different imaging approaches, starting with confocal fluorescence microscope and 2D-STED approaches while holding a chromosome with two optically trapped microspheres, and ultimately aiming for 3D-STED imaging of chromosomes held with four microspheres, using the newly developed instrument of task 1.2. We envision that this super-resolution imaging of different kinds provided by UCPH and UCL will result in deep insights in chromosome structure.