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Biophys Rep. 2018;4(4):215-221. doi: 10.1007/s41048-018-0065-z. Epub 2018 Aug 31.

Hessian single-molecule localization microscopy using sCMOS camera.

Xue F1,2, He W1, Xu F3, Zhang M1, Chen L4, Xu P1,2.

Author information

1Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China.
2College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049 China.
3Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907 USA.
4State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, 100871 China.


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 sCMOS camera can effectively increase the imaging rate due to its large field of view and fast imaging speed. Using an sCMOS camera for SMLM imaging can significantly improve the imaging time resolution, but the unique single-pixel-dependent readout noise of sCMOS 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 pixel-dependent readout noise of sCMOS 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.


Hessian; SMLM; Super-resolution; sCMOS

Conflict of interest statement

Fudong Xue, Wenting He, Fan Xu, Mingshu Zhang, Liangyi Chen, and Pingyong Xu declare that they have no conflict of interest.This article does not contain any studies with human or animal subjects performed by any of the authors.

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