In this webinar we presented a recently developed GPU-enabled high-speed analytical super-resolution approach – Super-Resolution Radial Fluctuations (SRRF, reads as ‘surf’). Compared to PALM and STORM, the requirement for sparse activation of fluorophores is circumvented in SRRF by calculating the temporal correlations present after applying a simple and fast image transform that quantifies the radial symmetries in short image sequences.
The applicability of SRRF to non-photoswitching datasets makes super-resolution possible with illumination orders of magnitude lower than methods such as SMLM or STED. It also enables live-cell imaging with conventional fluorophores using modern widefield, confocal or TIRF microscopes achieving resolutions better than 150 nm at 1 frame per second.
We also examined the basis of SRRF and demonstrate its capacity for live-cell over timescales of several minutes with sufficiently low photo-toxicity to allow normal cell behavior.
Andor have recently released a new implementation of SRRF, called SRRF-Stream, offering the capability to perform super-resolution in Real Time. SRRF-Stream processes data at up to 30x faster than the corresponding ImageJ post processing implementation of SRRF (NanoJ-SRRF). This furthermore permits image acquisition and SRRF processing to happen in parallel, resulting in a massive overall workflow improvement.
SRRF-Stream is available for iXon Ultra and iXon Life EMCCD cameras, accessible initially to users of either MicroManager software or the Andor SDK. Through uManager, the same acquisition control panel is maintained and super-resolution images are output rather than conventional images, either in time-series, mutli-channel or z-stack. SRRF-Stream processes data much faster than the iXon can acquire data, meaning that SRRF can even be operated in Live Mode.
Category: Webinar Recording