Quantitative high spatiotemporal imaging of biological processes

Super-resolution microscopy has revolutionized fluorescence imaging providing access to length scales that are much below the diffraction limit. The super-resolution methods have the potential for novel discoveries in biology. However, certain technical limitations must be overcome for this potential to be fulfilled. One of the main challenges is the use of super-resolution to study dynamic events in living cells. In addition, the ability to extract quantitative information from the super-resolution images is confounded by the complex photophysics that the fluorescent probes exhibit during the imaging. Here, we will review recent developments we have been implementing to overcome these challenges and introduce new steps in automated data acquisition towards high-throughput imaging.