3D tomography of cells in micro-channels
3D tomography of cells in micro-channels
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DOI：
10.1063/1.4986392
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发表时间：
2017-09-04
影响因子：
4
通讯作者：
Wagner, C.
中科院分区：
文献类型：
--
作者：
Quint, S.;Christ, A. F.;Wagner, C.
我们结合联合收割机共聚焦成像、微流体和图像分析来记录流动中细胞的3D图像。这使我们能够每分钟恢复数百个活细胞的完整3D表示。而3D共焦成像到目前为止仅限于稳定的标本，我们克服了这一限制，并提出了一种方法来访问移动物体的3D形状。我们的原则的关键是微通道相对于显微镜的焦平面的倾斜布置。这迫使细胞以倾斜的方式穿过焦平面。因此，记录通过细胞的各个层，然后可以将其组装以获得体积表示。完整的3D信息允许与理论和数值预测进行详细比较，例如，2D成像。我们的技术是通过研究在微通道中流动的红细胞来体现的，反映了微血管系统中的主要条件。我们观察到两种非常不同的形状：“羊角面包”和“拖鞋”。“此外，我们还对实验进行了3D数值模拟，以确认观察结果。由于流动中细胞的3D共聚焦成像尚未实现，因此我们在流式细胞术领域看到了很高的潜力，其中细胞分类迄今为止主要依赖于1D散射和荧光信号。由AIP出版社出版。
We combine confocal imaging, microfluidics, and image analysis to record 3D-images of cells in flow. This enables us to recover the full 3D representation of several hundred living cells per minute. Whereas 3D confocal imaging has thus far been limited to steady specimens, we overcome this restriction and present a method to access the 3D shape of moving objects. The key of our principle is a tilted arrangement of the micro-channel with respect to the focal plane of the microscope. This forces cells to traverse the focal plane in an inclined manner. As a consequence, individual layers of passing cells are recorded, which can then be assembled to obtain the volumetric representation. The full 3D information allows for a detailed comparison with theoretical and numerical predictions unfeasible with, e.g., 2D imaging. Our technique is exemplified by studying flowing red blood cells in a micro-channel reflecting the conditions prevailing in the microvasculature. We observe two very different types of shapes: "croissants" and "slippers." Additionally, we perform 3D numerical simulations of our experiment to confirm the observations. Since 3D confocal imaging of cells in flow has not yet been realized, we see high potential in the field of flow cytometry where cell classification thus far mostly relies on 1D scattering and fluorescence signals. Published by AIP Publishing.