Hessian single-molecule localization microscopy using sCMOS camera

Single-molecule localization microscopy(SMLM) has the highest spatial resolution among the existing super-resolution imaging techniques, but its temporal resolution needs further improvement. An s CMOS camera can effectively increase the imaging rate due to its large field of view and fast imaging speed. Using an s CMOS camera for SMLM imaging can significantly improve the imaging time resolution, but the unique single-pixel-dependent readout noise of s CMOS cameras severely limits their application in SMLM imaging. This paper develops a Hessian-based SMLM(Hessian-SMLM) method that can correct the variance, gain, and offset of a single pixel of a camera and effectively eliminate the pixeldependent readout noise of s CMOS cameras, especially when the signal-to-noise ratio is low. Using Hessian-SMLM to image mEos3.2-labeled actin was able to significantly reduce the artifacts due to camera noise.