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Single-Molecule Localization Microscopy with sCMOS Cameras

机译:带有sCMOS相机的单分子定位显微镜

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摘要

The ability of optical superresolution microscopy to circumvent the so-called "diffraction limit" has revolutionized the use of fluorescence microscopy to study subcellular and molecular-scale biological processes. Single-molecule localization microscopy (SMLM)-also commonly referred to as PALM, STORM and dSTORM, among other acronyms-overcomes the resolution limit by measuring the position of large numbers of marker molecules and can achieve effective lateral detail resolution in excess of 20 nm~(1-4) in addition to similar improvements in axial resolution. Typically, to reconstruct a single 2-D or 3-D image, several thousand camera frames have to be acquired and analyzed.~5 Preferred camera technologies combine the ability to sustain high frame rates with the capability to detect the comparatively low light levels associated with single-molecule fluorescence emission. Such cameras should enable the rapid image generation that often is required for live-cell recordings.
机译:光学超分辨率显微镜规避所谓的“衍射极限”的能力彻底改变了荧光显微镜用于研究亚细胞和分子规模生物学过程的能力。单分子定位显微镜(SMLM)-通常也称为PALM,STORM和dSTORM,以及其他首字母缩略词-通过测量大量标记分子的位置来克服分辨率极限,并且可以实现超过20 nm的有效横向细节分辨率除了轴向分辨率方面的类似改进外,还可以达到〜(1-4)。通常,要重建单个2D或3D图像,必须获取并分析数千个摄像机帧。〜5首选摄像机技术将维持高帧速率的能力与检测相关的相对较低的光照水平的能力结合在一起具有单分子荧光发射。这样的摄像机应该能够快速生成图像,这通常是活细胞录制所必需的。

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