Epi-illumination gradient light interference microscopy for imaging opaque structures
Epi-illumination gradient light interference microscopy for imaging opaque structures
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DOI：
10.1038/s41467-019-12634-3
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发表时间：
2019-10-16
影响因子：
16.6
通讯作者：
Popescu, Gabriel
中科院分区：
文献类型：
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作者：
Kandel, Mikhail E.;Hu, Chenfei;Popescu, Gabriel
多重散射和吸收限制了光对生物组织成像的深度。在厚的未标记的标本中，多次散射使场的相位随机化，而吸收则使长光路传播的光衰减。这些障碍限制了透射成像的性能。为了缓解这些挑战，我们开发了一种外延照度梯度光干涉显微镜（epi-GLIM），作为一种适用于散装或不透明样品的无标签相位成像方式。Epi-GLIM可以研究数百微米厚的浑浊结构，否则就无法透射光。我们用各种与透射几何成像不相容的人造和生物样品证明了这种方法：半导体晶片、不透明和双折射基底上的样品、微孔板上的细胞和大块组织。我们证明了epi-GLIM数据可以用于解决反散射问题和重建单细胞和模式生物的断层扫描。
Multiple scattering and absorption limit the depth at which biological tissues can be imaged with light. In thick unlabeled specimens, multiple scattering randomizes the phase of the field and absorption attenuates light that travels long optical paths. These obstacles limit the performance of transmission imaging. To mitigate these challenges, we developed an epi-illumination gradient light interference microscope (epi-GLIM) as a label-free phase imaging modality applicable to bulk or opaque samples. Epi-GLIM enables studying turbid structures that are hundreds of microns thick and otherwise opaque to transmitted light. We demonstrate this approach with a variety of man-made and biological samples that are incompatible with imaging in a transmission geometry: semiconductors wafers, specimens on opaque and birefringent substrates, cells in microplates, and bulk tissues. We demonstrate that the epi-GLIM data can be used to solve the inverse scattering problem and reconstruct the tomography of single cells and model organisms.