Optimal acquisition scheme for subwavelength localization microscopy of bleachable fluorophores.

We consider the problem of optimizing superresolution microscopy with photoswitchable molecules that irreversibly photobleach. Using variational methods, we show that the number of single-molecule images is maximized in a simple scheme with a constant number of activated fluorophores per cycle. For high-speed acquisition, deviations from the optimal scheme do reduce the information collected but fortuitously also reduce certain types of errors, making the scheme very robust. Finally, we calculate the amount of information lost due to bleaching and noise.