2025年8月19日发布
活细胞单分子荧光成像方法在各个生命科学研究领域发挥着重要作用,而探测器的灵敏度和分辨率对于成像性能至关重要。
为了达到最佳单分子成像性能,Chinese Academy of Sciences下属Institute of Genetics and Developmental Biology的 Dr. Jiachao Xu搭建了TIRF显微镜,并在最近几年不断改进。
近期,Dr. Xu在其活细胞血管紧张素II 1型受体 (AT1R) 单分子荧光成像实验中,比较了EM-CCD与qCMOS两种相机的性能。
Jiachao Xu 博士的实验要求探测器具有极低噪声,以便从相对干净的背景中轻松识别单分子荧光。
根据他的经验,EM-CCD 可以提供良好的信噪比。之前他也尝试过一些其他类型的相机,但没有一台能达到类似的性能。
但这次 qCMOS 表现出色,即使在快速模式 (0.43e r.m.s.) 下读出噪声也几乎与 EM-CCD 一样低。从 qCMOS 和 EM-CCD 采集的图像中可以观察到几乎相同数量的荧光单分子。
此外,qCMOS 具有 4.6 ㎛ 的更小像素尺寸,因而可根据成像需求在某些模式下使用:4.6 ㎛(1×1 像素合并)、9.2 ㎛(2×2 像素合并)和 18.4 ㎛(4×4 像素合并)。例如,为了平衡灵敏度和定位精度,Xu 博士选择了 9.2 ㎛(2×2 像素合并)设置以获得最佳性能。
综上所述,qCMOS 在单分子荧光成像方面表现出与 EM-CCD 相当甚至更好的性能。其灵敏度、速度和分辨率优势可为不同科学成像用途提供更多选择。
样本:AT1R-EYFP
曝光时间:100 ms
光学系统:TIRF
物镜: 100× NA 1.49
激光器:488nm 1mW
Dr. Jiachao Xu is from Dr. Kangmin He’s laboratory. Their research is highly praised in the field. The research method of real-time live-cell single particle tracking using TIRFM is "exciting, which will promote many peers to carry out similar research".
*本页显示的内容为采访时的可用信息。
ORCA-Quest 2 是一款新的 qCMOS® 相机,也是 ORCA-Quest 的后续产品,具有多项深化进步,例如在极低噪声扫描模式下读出速度更快,紫外区域灵敏度更高。
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