Three-dimensional, Portable, Inexpensive, and Reusable Tomographic Microscopy

项目摘要 三维（3D）显微镜为生物学研究和医学研究提供了许多希望。 应用.然而，目前仅限于资源充足的实验室， 基础设施演进许多3D显微技术（例如，共焦）依赖于将光聚焦在一个小的 这需要昂贵的专用设备。光学投影 断层扫描（OPT）是一种3D成像技术，利用传统的显微镜设备;而不是 OPT将光聚焦在特定位置，从多个角度对样品进行成像，以重建3D体积。 这是一种非常有效的方法，用于小型半透明物体（例如，小鼠胎儿、寄生虫和 大细菌）。虽然OPT可能是一种成本较低的3D显微镜方法，但现有系统 仍然昂贵和庞大。我们建议利用无处不在的高质量计算 和成像硬件，使OPT设备，这是廉价的， 移植的.我们将创建一个智能手机扩展，它可以对旋转样本进行鲁棒成像，并使用 手机的计算硬件来重建3D体积。将采用两种成像模式： 可见波段衰减显微术（例如，明场）和发光显微术（例如，生物发光的）。 智能手机扩展的组件可以是3D打印的，激光切割的丙烯酸，也可以是商业上容易的。 available.因此，制造该装置的成本非常小，并且装置的总重量非常小。 非常低;所有组件的总成本将低于50美元。扩展的组件很容易 在现场组装，允许该装置在小包装中运输。由于其低成本和 尺寸，OPT显微镜也可以作为一个有用的工具，用于教育目的（例如，在一项 涉及光学的本科生实验课程）和用于生成用于层析成像算法的真实的数据 发展为了验证该设备，我们将建立两个具有三维特征的幻影， 我们评估调制传递函数：一个用于衰减显微镜，另一个用于发光显微镜。 显微镜目标1：构建智能手机的可见波段断层扫描显微镜扩展。目标1A： 根据约10 mm样本的可见波段正弦图数据实现3D锥束重建 分辨率约为10 µm。目标1B：使用多镜头光学系统提高图像分辨率 系统目的2：通过适当修改生物发光断层成像显微镜， 层析重建算法目标3：构建两个具有尺寸特征的货架稳定的3D体模 从1 μm到200 μm不等。

PROJECT SUMMARY Three-dimensional (3D) microscopy offers many promises for biological investigations and medical applications. However, it is currently limited to well-resourced laboratories in settings with established infrastructure. Many 3D microscopy techniques (e.g., confocal) rely on focusing light at an array of small locations within the sample, which requires expensive and specialized equipment. Optical Projection Tomography (OPT) is a 3D imaging technique that utilizes traditional microscopy equipment; instead of focusing the light at specific locations, OPT images a sample from many angles to reconstruct a 3D volume. It is a very effective method for 3D imaging of small translucent objects (e.g., mouse fetuses, parasites, and large bacteria). While it is possible for OPT to be a lower-cost method of 3D microscopy, existing systems remain expensive and large. We propose to take advantage of the ubiquitous and high-quality computing and imaging hardware available in smartphones to make an OPT device that is inexpensive and extremely portable. We will create a smartphone extension that robustly images a rotating sample and uses the phone’s computational hardware to reconstruct the 3D volume. Two imaging modalities will be pursued: visible-band attenuation microscopy (e.g., brightfield) and luminescent microscopy (e.g., bioluminescent). The components of the smartphone extension are either 3D printed, laser cut acrylic, or readily commercially available. Thus, the cost of manufacturing the device is extremely small and the total weight of the device is very low; the total cost of all components will be less than $50. The components of the extension are easily assembled on site, permitting the device to be transported with in a small package. Due to its low cost and size, the OPT microscope can also serve as a useful tool for educational purposes (e.g., as part of an undergraduate laboratory course involving optics) and for generating real data for tomographic algorithm development. To validate the device, we will build two phantoms with three-dimensional features that allow us to evaluate the Modulation Transfer Function: one for attenuation microscopy and another for luminescent microscopy. Aim 1: Build a visible-band tomographic microscope extension to a smartphone. Aim 1A: Implement 3D cone-beam reconstruction from visible-band sinogram data for samples approximately 10 mm in size with approximately 10 µm resolution. Aim 1B: Improve the image resolution with a multi-lens optical system. Aim 2: Implement bioluminescent tomographic microscopy by appropriately modifying the tomographic reconstruction algorithm. Aim 3: Build two shelf-stable 3D phantoms with features of sizes varying from 1 µm to 200 µm.