Super-resolution localization microscopy (SRLM) has been an important tool for biology because it brings the spatial resolution of optical microscopy down to nanoscale through a relative simple setup. The selection of a suitable low-light camera is undoubtedly critical in SRLM. Conventionally, Electron Multiplying Charge Coupled Device (EMCCD) cameras are used to detect the weak fluorescence from single molecules, mainly because this type of low-light cameras provides not only ultra-low read noise at high readout rate, but also ultimate quantum efficiency. Starting from 2009, scientific Complementary Metal Oxide Semiconductor (sCMOS) cameras has been actively explored as an alternative to the popular EMCCD cameras in SRLM. sCMOS cameras provide simultaneously low read noise, high readout speed, and large pixel array; however, the relatively low quantum efficiency of sCMOS cameras has been a major limitation for its wide-spread use in SRLM. In this talk, we will evaluate the imaging performance of a back-illuminated sCMOS camera (called Dhyana 95 from Tucsen) which is claimed to be the world’s first 95% QE sCMOS camera. The evaluation is based on a new methodology which is designed to provide paired images from two tested cameras under almost identical experimental conditions. We compare this 95% QE sCMOS camera with a representative front-illuminated sCMOS camera and a popular back-illuminated EMCCD camera. We believe that this study can provide useful information for selecting a suitable low-light detector for SRLM.
Zhenli Huang, Yujie Wang, and Lingxi Zhao, "Quantitative performance evaluation of a back-illuminated sCMOS camera with 95% QE for super-resolution localization microscopy (Conference Presentation)," Proc. SPIE 10500, Single Molecule Spectroscopy and Superresolution Imaging XI, 105000A (Presented at SPIE BiOS: January 27, 2018; Published: 14 March 2018); https://doi.org/10.1117/12.2286840.5751643368001.
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