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Wide-field high-resolution 3D microscopy with Fourier ptychographic diffraction tomography
Optics and Lasers in Engineering ( IF 3.5 ) Pub Date : 2020-05-01 , DOI: 10.1016/j.optlaseng.2020.106003
Chao Zuo , Jiasong Sun , Jiaji Li , Anand Asundi , Qian Chen

Abstract We report a computational 3D microscopy technique, termed Fourier ptychographic diffraction tomography (FPDT), that iteratively stitches together numerous variably illuminated, low-resolution images acquired with a low-numerical aperture (NA) objective in 3D Fourier space to create a wide field-of-view (FOV), high-resolution, depth-resolved complex refractive index (RI) image across large volumes. Unlike conventional optical diffraction tomography (ODT) approaches that rely on controlled bright-field illumination, holographic phase measurement, and high-NA objective detection, FPDT employs tomographic RI reconstruction from low-NA intensity-only measurements. In addition, FPDT incorporates high-angle dark-field illuminations beyond the NA of the objective, significantly expanding the accessible object frequency. With FPDT, we present the highest-throughput ODT results with 390 nm lateral resolution and 899 nm axial resolution across a 10 × FOV of 1.77 mm2 and a depth of focus of ~ 20 µm. Billion-voxel 3D tomographic imaging results of biological samples establish FPDT as a powerful non-invasive and label-free tool for high-throughput 3D microscopy applications.

中文翻译:

具有傅里叶 ptychographic 衍射断层扫描的宽视场高分辨率 3D 显微镜

摘要 我们报告了一种计算 3D 显微技术,称为傅立叶 ptychographic 衍射断层扫描 (FPDT),它迭代地将在 3D 傅立叶空间中使用低数值孔径 (NA) 物镜获取的许多可变照明的低分辨率图像拼接在一起,以创建宽视场大体积的视野 (FOV)、高分辨率、深度分辨复折射率 (RI) 图像。与依赖受控明场照明、全息相位测量和高 NA 物镜检测的传统光学衍射断层扫描 (ODT) 方法不同,FPDT 采用仅低 NA 强度测量的断层 RI 重建。此外,FPDT 结合了超出物镜 NA 的高角度暗场照明,显着扩大了可接近的物体频率。使用 FPDT,我们展示了最高通量的 ODT 结果,横向分辨率为 390 nm,轴向分辨率为 899 nm,10 × FOV 为 1.77 mm2,焦深约为 20 µm。生物样本的 10 亿体素 3D 断层成像结果使 FPDT 成为用于高通量 3D 显微应用的强大无创和无标记工具。
更新日期:2020-05-01
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