Ma Ruyu, Santino Luciano M, Chobola Tomáš, Armbrust Niklas, Geilenkeuser Julian, Sukumaran Sapthagiri, Jing Zhizi, Levkina Anastasia, Ridderbeek Korneel, Peng Tingying, Truong Dong-Jiunn Jeffery, Doll Sebastian, Westmeyer Gil Gregor, Cui Jian
Helmholtz Pioneer Campus, Helmholtz Munich, Neuherberg, Germany.
Helmholtz AI, Helmholtz Munich, Neuherberg, Germany.
Nat Methods. 2025 May 29. doi: 10.1038/s41592-025-02694-3.
Bioluminescence is an attractive alternative to fluorescence for live-cell imaging; however, its low intensity has prevented widespread adoption. Specialized microscopes compensate by sacrificing spatial resolution, field of view and dynamic range-constraints imposed by the highest-sensitivity camera to date: the electron-multiplying charge-coupled device. Recently, quanta image sensor (QIS) technology has emerged for low-light imaging. Here, we show that a commercial QIS camera has exceptional sensitivity; however, its sensor dimensions necessitate a microscope designed to maximize its properties. We introduce a Keplerian-telescope-inspired microscope setup that, with the QIS, results in modestly improved signal-to-noise ratios at substantially higher spatial resolution, field of view and dynamic range, relative to the state of the art. The telescopic design also confers modularity, enabling multimodal imaging with epifluorescence. The 'QIScope' makes bioluminescence a viable tool for technically challenging live-cell experiments such as monitoring intracellular and extracellular vesicles simultaneously and the dynamics of low-abundance proteins.
生物发光作为一种用于活细胞成像的方法，是荧光成像颇具吸引力的替代方案；然而，其低强度阻碍了它的广泛应用。专门的显微镜通过牺牲空间分辨率、视野和动态范围来进行补偿，这些限制是由迄今为止最高灵敏度的相机——电子倍增电荷耦合器件所施加的。最近，量子图像传感器（QIS）技术已出现用于低光成像。在此，我们展示了一款商用QIS相机具有卓越的灵敏度；然而，其传感器尺寸需要设计一种能使其性能最大化的显微镜。我们引入了一种受开普勒望远镜启发的显微镜设置，该设置与QIS相结合，相对于现有技术，在显著更高的空间分辨率、视野和动态范围下，能适度提高信噪比。这种望远镜式设计还赋予了模块化特性，能够实现落射荧光的多模态成像。“QIScope”使生物发光成为用于技术上具有挑战性的活细胞实验的可行工具，比如同时监测细胞内和细胞外囊泡以及低丰度蛋白质的动态变化。
