Huang Zhen-Li, Zhu Hongyu, Long Fan, Ma Hongqiang, Qin Lingsong, Liu Yongfeng, Ding Jiuping, Zhang Zhihong, Luo Qingming, Zeng Shaoqun
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074, China.
Opt Express. 2011 Sep 26;19(20):19156-68. doi: 10.1364/OE.19.019156.
In the community of localization-based super-resolution microscopy (or called localization microscopy), it is generally believed that the emission of single molecules is so weak that an EMCCD (electron multiplying charge coupled device) camera is necessary to be used as the detector by eliminating read noise. Here we evaluate the possibility of a new kind of low light detector, scientific complementary metal-oxide-semiconductor (sCMOS) camera in localization microscopy. We demonstrate experimentally that sCMOS is capable of imaging actin bundles with FWHM diameter of 37 nm, evidencing the capability of sCMOS in localization microscopy. We further characterize the noise performance of sCMOS and find out that, with the use of a bright fluorescence probe such as d2EosFP, localization microscopy imaging is now working in the shot noise limited region.
在基于定位的超分辨率显微镜（或称为定位显微镜）领域，人们普遍认为单分子的发射非常微弱，以至于需要使用电子倍增电荷耦合器件（EMCCD）相机作为探测器来消除读取噪声。在此，我们评估了一种新型低光探测器——科学互补金属氧化物半导体（sCMOS）相机在定位显微镜中的应用可能性。我们通过实验证明，sCMOS能够对半高宽直径为37 nm的肌动蛋白束进行成像，这证明了sCMOS在定位显微镜中的能力。我们进一步对sCMOS的噪声性能进行了表征，发现使用如d2EosFP这样的明亮荧光探针时，定位显微镜成像目前处于散粒噪声限制区域。
