Prof. Pingyong Xu
Dr. Mingshu Zhang
Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences
Single-molecule localization microscopy (SMLM) techniques, such as photoactivation localization microscopy (PALM), achieve some of the highest spatial resolutions among all the super-resolution imaging methods. However, image reconstruction in PALM requires a large number of raw images, which leads to low temporal resolution. Limited photo-controllable fluorescent protein probes (PCFPs) also restrict the widespread application of SMLM.
Prof. Pingyong Xu’s lab is engaged in the development of new super-resolution microscopy technologies. Previously, his group developed a new super-resolution method named single-molecule-guided Bayesian localization microscopy (SIMBA). It uses only 100 raw frames to calculate a super-resolution image with a 50 nm spatial resolution, which efficiently increases the temporal resolution of SMLM.
To achieve higher temporal resolution with SIMBA, prof. Xu intended to develop imaging techniques that improved on SIMBA and required even less raw frames for reconstruction. Another way to improve temporal resolution is to use a camera with a higher frame rate, with sufficient sensitivity to image with shorter exposure times. However, the slow readout speed of a previous EMCCD camera solution and the suboptimal properties of existing PCFPs hindered them from further increasing the temporal resolution of SIMBA.
[We] benefited from the high speed and sensitivity of Prime 95B, we could further increase the temporal resolution of SIMBA with shorter exposure times.