Strasser Franziska, Offterdinger Martin, Piestun Rafael, Jesacher Alexander
Division of Biomedical Physics, Medical University of Innsbruck, Müllerstraße 44, 6020 Innsbruck, Austria.
Division of Neurobiochemistry, Biooptics, Medical University of Innsbruck, Innrain 80-82, 6020 Innsbruck, Austria.
Biomed Opt Express. 2019 Apr 22;10(5):2513-2527. doi: 10.1364/BOE.10.002513. eCollection 2019 May 1.
For decades, the confocal microscope has represented one of the dominant imaging systems in biomedical imaging at sub-cellular lengthscales. Recently, however, it has increasingly been replaced by a related, but more powerful successor technique termed image scanning microscopy (ISM). In this article, we present ISM capable of measuring spectroscopic information such as that contained in fluorescence or Raman images. Compared to established confocal spectroscopic imaging systems, our implementation offers similar spectral resolution, but higher spatial resolution and detection efficiency. Color sensitivity is achieved by a grating placed in the detection path in conjunction with a camera collecting both spatial and spectral information. The multidimensional data is processed using multi-view maximum likelihood image reconstruction. Our findings are supported by numerical simulations and experiments on micro beads and double-stained HeLa cells.
几十年来，共聚焦显微镜一直是生物医学成像中在亚细胞长度尺度上占主导地位的成像系统之一。然而，最近它越来越多地被一种相关但更强大的后续技术——图像扫描显微镜（ISM）所取代。在本文中，我们展示了能够测量光谱信息（如荧光或拉曼图像中包含的信息）的ISM。与已有的共聚焦光谱成像系统相比，我们的实现方式具有相似的光谱分辨率，但空间分辨率和检测效率更高。通过在检测路径中放置一个光栅并结合收集空间和光谱信息的相机来实现颜色敏感度。使用多视图最大似然图像重建来处理多维数据。我们的发现得到了对微珠和双重染色的HeLa细胞的数值模拟和实验的支持。
