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）间接成像提议通过SBIR资助机制，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。荧光探针、探测器技术和微加工技术的最新进展使构建图像映射光谱仪（IMS）成为可能，这是一种快速、实时定量光谱成像的设备。IMS是一种宽视场方法，可以同时从每个像素获取全光谱信息。与扫描高光谱系统相比，它具有优越的信噪比，可以与光学切片方法（如Nipkow磁盘）一起使用。IMS通过空间重定向图像区域来获取线之间的空间，并使用多棱镜元素同时获取目标的光谱和空间信息。通过将CCD 2D图像传感器的像素位置重新映射到各自的体素（x, y,;），重建最终的光谱立方体。这是一个第一阶段的提案，其中我们将专注于(1)开发一个更大格式的IMS系统，能够收集大小为500 x 500 x 48的（x, y,;）数据立方体，初始波长范围为450到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。与此同时，该项目将追求(2)开发以最低成本制造图像映射器的方法-制造过程目前昂贵且耗时，根据尺寸和复杂性，每个部件需要100多个小时。我们将追求一种新的钻石加工方法，这种方法有可能大大缩短加工时间。此外，我们将实施(3)自动校准程序和软件，用于实时数据分析和可视化，从而优化性能，提高分辨率和帧率光谱解混能力。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。综上所述，IMS通过减少光毒性和光漂白以及允许在高帧率下进行高光谱分析，具有显著推进细胞成像领域广泛应用的潜力。为了进一步扩大其影响，在未来，我们计划将IMS与光学切片结合使用结构化照明，Nipkow盘共聚焦和/或空间反卷积。这些四维成像系统（X， Y， Z,;）将进一步提高采集图像的信噪比，提高采集速度。

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. PUBLIC HEALTH RELEVANCE: The project targets the development of a modern spectrometer called high sampling Image Mapping Spectrometer enabling high resolution spectral imaging in real time. In consequence researchers will be able to rapidly advance the investigation of live cells with multiple fluorescent contrasts. The instrument's principle allows obtaining spectral information for entire image without scanning and thus improve signal to noise ratio and limit photo-bleaching effects. It also allows more efficient investigation of transient biological events. Technologies applied in the project and their low cost may potentially allow access of larger group of scientists to spectral imaging instrumentation.