White-light diffraction tomography of unlabelled live cells
White-light diffraction tomography of unlabelled live cells
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DOI：
10.1038/nphoton.2013.350
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发表时间：
2014-03-01
期刊：
影响因子：
35
通讯作者：
Popescu, Gabriel
中科院分区：
文献类型：
--
作者：
Kim, Taewoo;Zhou, Renjie;Popescu, Gabriel
我们提出了一种称为白光衍射层析成像(WDT)的技术，用于对活的未标记细胞等微观透明对象进行成像。该方法将衍射层析成像扩展到白光照明和成像，而不是散射面测量。我们的实验是使用传统的相差显微镜进行的，该显微镜升级了一个模块来测量定量的相位图像。通过扫描焦点穿过物体并获取一叠相分辨图像来重建物体的轴向尺寸。我们重建了活的、未标记的红细胞的三维结构，并将结果与共聚焦和扫描电子显微镜图像进行了比较。350 nm的横向分辨率和900 nm的轴向分辨率在高分辨率下揭示了大肠杆菌细胞中的亚细胞结构。这一结果确立了WDT作为一种以非侵入性和无标记的方式测量三维亚细胞结构的手段。
We present a technique called white-light diffraction tomography (WDT) for imaging microscopic transparent objects such as live unlabelled cells. The approach extends diffraction tomography to white-light illumination and imaging rather than scattering plane measurements. Our experiments were performed using a conventional phase contrast microscope upgraded with a module to measure quantitative phase images. The axial dimension of the object was reconstructed by scanning the focus through the object and acquiring a stack of phase-resolved images. We reconstructed the three-dimensional structures of live, unlabelled, red blood cells and compared the results with confocal and scanning electron microscopy images. The 350 nm transverse and 900 nm axial resolution achieved reveals subcellular structures at high resolution in Escherichia coli cells. The results establish WDT as a means for measuring three-dimensional subcellular structures in a non-invasive and label-free manner.