High-speed in vitro intensity diffraction tomography
High-speed in vitro intensity diffraction tomography
复制标题
DOI：
10.1117/1.ap.1.6.066004
复制
发表时间：
2019-11-01
影响因子：
17.3
通讯作者：
Tian, Lei
中科院分区：
文献类型：
--
作者：
Li, Jiaji;Matlock, Alex;Tian, Lei
我们展示了一种利用环形照明(AIDT)的无标记、无扫描的强度衍射层析成像技术来快速表征体外大体积三维(3-D)折射率分布。通过将照明几何形状与显微镜的光瞳进行最佳匹配，我们的技术将数据需求减少了60倍，从而实现了高速10赫兹的体积速率。利用8幅强度图像，恢复了约350微米×100微米×20微米的体积，横向近衍射限分辨率为487 nm，轴向分辨率为3.4微米，所获得的大体积比和高分辨率使复杂生物样品的三维定量相位成像可以跨多个长度尺度进行。我们展示了AIDT在单细胞硅藻微藻、具有本地细菌的口腔上皮细胞群和秀丽隐杆线虫活体标本上的能力。在这些样本中，我们恢复了具有最小运动伪影的宏观细胞结构、亚细胞细胞器和动态微生物组织。量化这些特征在肿瘤学、免疫学和细胞病理生理学中具有重要的实用价值，在这些领域中，这些形态特征被评估为疾病、寄生虫和新药治疗的存在的变化。最后，我们对AIDT系统进行了仿真，以突出所提出的技术的准确性和敏感性。AIDT显示出作为一种强大的高速、无标记计算显微镜方法的前景，适用于需要自然成像来实时评估对样本的环境影响的应用。
We demonstrate a label-free, scan-free intensity diffraction tomography technique utilizing annular illumination (aIDT) to rapidly characterize large-volume three-dimensional (3-D) refractive index distributions in vitro. By optimally matching the illumination geometry to the microscope pupil, our technique reduces the data requirement by 60 times to achieve high-speed 10-Hz volume rates. Using eight intensity images, we recover volumes of similar to 350 mu m x 100 mu m x 20 mu m, with near diffraction-limited lateral resolution of similar to 487 nm and axial resolution of similar to 3.4 mu m. The attained large volume rate and high-resolution enable 3-D quantitative phase imaging of complex living biological samples across multiple length scales. We demonstrate aIDT's capabilities on unicellular diatom microalgae, epithelial buccal cell clusters with native bacteria, and live Caenorhabditis elegans specimens. Within these samples, we recover macroscale cellular structures, subcellular organelles, and dynamic micro-organism tissues with minimal motion artifacts. Quantifying such features has significant utility in oncology, immunology, and cellular pathophysiology, where these morphological features are evaluated for changes in the presence of disease, parasites, and new drug treatments. Finally, we simulate the aIDT system to highlight the accuracy and sensitivity of the proposed technique. aIDT shows promise as a powerful high-speed, label-free computational microscopy approach for applications where natural imaging is required to evaluate environmental effects on a sample in real time.