Scipioni, L. et al. Nat. Commun. 9, 5120 (2018).

Fluorescence fluctuation techniques are established approaches for studying dynamic behaviors of labeled molecules within cells. However, various types of fluorescence fluctuation analyses have different acquisition parameters, which makes simultaneous acquisition challenging. Scipioni et al. have developed comprehensive correlation analysis, which allows many advanced fluorescence fluctuation techniques to be implemented in a single, simultaneous analysis. Their approach is based on the ultrafast Airyscan detector in the Zeiss LSM 880, which can operate at around 1 million frames per second. The researchers show that with this instrument one can study both fluorescently labeled probes and the environment in which they are diffusing, via the simultaneous acquisition of parameters such as oligomerization state, diffusion coefficient, diffusion modality, and barriers to diffusion. They demonstrated their approach by mapping GFP diffusion in cells.