Optimization of sample, imaging and data processing parameters is an essential task in localization based super-resolution microscopy, where the final image quality strongly depends on the imaging of single isolated fluorescent molecules. A computational solution that uses a simulator software for the generation of test data stacks was proposed, developed and tested. The implemented advanced physical models such as scalar and vector based point spread functions, polarization sensitive detection, drift, spectral crosstalk, structured background etc., made the simulation results more realistic and helped us interpret the final super-resolved images and distinguish between real structures and imaging artefacts.
Novák et al., Scientific Reports 7, Article number: 951 (2017) doi:10.1038/s41598-017-01122-7