Snapshot Image Mapping Spectrometer (IMS) for High Resolution Biological Imaging

描述(由申请人提供)：用于高分辨率生物成像的快照图像映射光谱仪(IMS)通过SBIR资助机制，建议开发一种创新的成像设备，该设备将允许经济的快照高光谱成像，用于实时显微镜和其他生物医学应用，并与大多数研究级光学显微镜兼容。在荧光探测器、探测器技术和微制造方面的最新进展现在使建造图像映射光谱仪(IMS)成为可能--一种用于快速、实时定量光谱成像的设备。IMS是一种从每个像素同时获取全光谱信息的宽域方法。与扫描高光谱系统相比，它具有更高的信噪比，并可与尼普科夫磁盘等光学切片方法一起使用。IMS的工作原理是在空间上重定向图像区域以获得线之间的空间，并使用多棱镜元件来同时获取关于对象的光谱和空间信息。通过将像素位置从CCD2D图像传感器重新映射到各自的体素(x，y，；)来重建最终的光谱立方体。这是第一阶段的提案，在这一阶段，我们将专注于(1)开发一个更大格式的IMS系统，能够采集大小为500 x 500 x 48的DataCube，初始波长范围为450至700 nm，并在几个活细胞成像应用中测试图像映射光谱仪与当前可用的光谱成像系统的对比。同时，该项目将致力于(2)开发以最低成本制造图像映射器的方法--目前制造过程昂贵且耗时，根据尺寸和复杂性的不同，每个部件需要100个小时。我们将寻求一种新的钻石规则制造方法，这种方法有可能大幅缩短制造时间。此外，我们还将实施(3)用于实时数据分析和可视化的自动校准程序和软件，从而优化性能、提高分辨率和帧速率光谱分解能力。这将首次为研究人员提供实时活细胞高光谱成像的即时、实时反馈。总之，IMS通过减少光毒性和光漂白，并允许在高帧速率下进行高光谱分析，有可能显著推进细胞成像领域的广泛应用。为了进一步扩大其影响，在未来，我们计划通过使用结构照明、尼普考圆盘共焦和/或空间去卷积将IMS与光学切片相结合。这些4维成像系统(X、Y、Z、；)将进一步提高采集图像的信噪比和速度。

DESCRIPTION (provided by applicant): Snapshot Image Mapping Spectrometer (IMS) for High Resolution Biological Imaging Indirect Imaging is proposing, through the SBIR funding mechanism, to develop an innovative imaging device that will allow economical snapshot hyperspectral imaging for real time microscopy and other biomedical applications, and is compatible with most research grade light microscopes. Recent advances in fluorescent probes, detector technology and micro-fabrication now make it possible to build an Image Mapping Spectrometer (IMS) - a device for rapid, real time quantitative spectral imaging. The IMS is a widefield method for acquiring full spectral information simultaneously from every pixel. It has superior signal-to-noise ratio compared to scanning hyperspectral systems and can be used with optical sectioning methods such as Nipkow disk. The IMS works by spatially redirecting image zones to obtain space between lines and using a multi-prism element to acquire simultaneously spectral and spatial information about the object. The final spectral cube is reconstructed by remapping the pixel locations from the CCD 2D image sensor to respective voxels (x, y, ;). This is a Phase I proposal, in which we will focus on (1) developing a larger format IMS system capable of collection a (x, y, ;) datacube of size 500 x 500 x 48 with an initial wavelength range of 450 to 700 nm and testing the Image Mapping Spectrometer against currently available spectral imaging systems in several live cell imaging applications. In parallel the project will pursue (2) developing the means to manufacture an Image Mapper at minimal costs - the fabrication process is currently expensive and time consuming taking 100+ hours/per part depending on the size and complexity. We will pursue a new diamond ruling fabrication approach that has a potential to dramatically shorten the fabrication time. In addition we will implement (3) automatic calibration procedures and software for real-time data analysis and visualization leading to optimized performance, improved resolution and frame-rate spectral unmixing capability. For the first time this will provide researchers with immediate, live feedback in real-time living cell hyperspectral imaging. In summary, the IMS has the potential to significantly advance a wide range of applications in the area of cellular imaging by reducing the phototoxicity and photobleaching and allowing hyperspectral analysis at high frame rates. To further its impact, in the future, we plan to combine the IMS with optical sectioning by using structured illumination, Nipkow disk confocal, and/or spatial deconvolution. These 4-dimensional imaging systems (X, Y, Z, ;) would further improve the signal-to-noise ratio of the collected images and improve their speed.