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

项目总结 三维(3D)显微镜为生物研究和医学提供了许多希望 申请。然而，它目前仅限于资源充足的实验室， 基础设施。许多3D显微镜技术(例如，共焦)依赖于将光聚焦在一系列小的 样本内的地点，这需要昂贵和专门的设备。光学投影 层析成像(OPT)是一种3D成像技术，它使用传统的显微镜设备；而不是 OPT将光线聚焦到特定位置，从多个角度对样本进行成像，以重建3D体积。 这是一种非常有效的方法，用于对小的半透明对象(如小鼠胚胎、寄生虫和 大型细菌)。虽然OPT有可能成为一种低成本的3D显微镜方法，但现有的系统 保持昂贵和庞大的规模。我们建议利用无处不在的高质量计算 和智能手机中可用的成像硬件，以制造廉价且极其 便携的。我们将创建一个智能手机扩展，它可以对旋转的样本进行健壮的图像处理，并使用 手机的计算硬件来重建3D体积。将采用两种成像方式： 可见波段衰减显微镜(例如，Brightfield)和发光显微镜(例如，生物发光)。 智能手机扩展模块的组件要么是3D打印的，要么是激光切割的亚克力，或者很容易商业化 可用。因此，制造该装置的成本非常小，并且该装置的总重量是 非常低；所有组件的总成本将不到50美元。扩展模块的组件很容易实现 现场组装，允许设备以小包装的形式运输。由于它的低成本和 此外，OPT显微镜还可用作教育用途的有用工具(例如，作为 涉及光学的本科实验室课程)，并用于为层析算法生成真实数据 发展。为了验证设备，我们将构建两个具有三维特征的幻影，以允许 美国将评估调制传递函数：一个用于衰减显微镜，另一个用于发光 显微镜。目标1：将可见波段断层扫描显微镜扩展到智能手机上。目标1A： 对约10 mm的样本实现可见波段正弦图数据的三维锥束重建 尺寸约10微米，分辨率约为10微米。目标1B：用多镜头光学系统提高图像分辨率 系统。目标2：通过适当修改 层析重建算法。目标3：建立两个具有大小特征的货架稳定的3D模型 从1微米到200微米不等。

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.