Hessian single-molecule localization microscopy using sCMOS camera.
Hessian single-molecule localization microscopy using sCMOS camera.
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DOI：
10.1007/s41048-018-0065-z
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发表时间：
2018
影响因子：
--
通讯作者：
Xu P
中科院分区：
文献类型：
--
作者：
Xue F;He W;Xu F;Zhang M;Chen L;Xu P
单分子定位显微镜（SMLM）在现有的超分辨率成像技术中具有最高的空间分辨率，但其时间分辨率有待进一步提高。 sCMOS相机由于视场大、成像速度快，可以有效提高成像速率。使用sCMOS相机进行SMLM成像可以显着提高成像时间分辨率，但sCMOS相机独特的单像素相关读出噪声严重限制了其在SMLM成像中的应用。本文提出了一种基于Hessian的SMLM（Hessian-SMLM）方法，可以校正相机单个像素的方差、增益和偏移，有效消除sCMOS相机的像素相关读出噪声，特别是在信噪比较低的情况下。使用 Hessian-SMLM 对 mEos3.2 标记的肌动蛋白进行成像能够显着减少由于相机噪声造成的伪影。
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.