使用极化正交断裂技术进行有丝分裂染色体的无标记显微镜检查
Label-free microscopy of mitotic chromosomes using the polarization Orthogonality Breaking technique
摘要：
目前可用于研究生物样品的绝大多数显微镜方法在成像之前需要染色。然而，在不同的情况下，以无标记的方式揭示特定细胞结构或细胞器的能力仍然是期望的。偏振显微镜一直被认为是一个有趣的替代荧光为基础的方法，以获得特异性的成像生物样品。在这项工作中，我们展示了如何一个原始的偏振成像技术，实现微波光子学，并被称为正交打破（OB）显微镜，可以提供信息偏振图像从一个快速和敏感的方式单次扫描的细胞样本。对于OB成像，用激光装置探测样品，同时产生频率偏移几十MHz的两个正交偏振。如果成像的样品显示出一些偏振特性，则两个偏振之间的正交性被破坏，导致可以用快速检测器检测到的拍音干涉信号。通过对不同细胞周期阶段的细胞系的OB偏振显微镜图像和荧光共聚焦图像的比较，发现在细胞分裂过程中致密染色体上存在内源性的偏振反差。该技术为有丝分裂染色体的无标记实时偏振成像铺平了道路，并在组织学和癌症诊断中具有进一步的潜在应用。
英文摘要：
The vast majority of the microscopy methods currently available to study biological samples require staining prior to imaging. Nevertheless, the ability to reveal specific cell structures or organelles in a label-free manner remains desirable in different contexts. Polarization microscopy has long been considered as an interesting alternative to fluorescence-based methods in order to gain specificity on the imaged biological samples. In this work, we show how an original polarization imaging technique, implementing micro-wave photonics and referred to as orthogonality-breaking (OB) microscopy, can provide informative polarization images from a single scan of the cell sample in a fast and sensitive way. For OB imaging, the sample is probed with a laser setup simultaneously generating two orthogonal polarizations shifted in frequency by a few tens of MHz. If the imaged samples display some polarimetric properties, the orthogonality between the two polarizations is broken, leading to a beatnote interference signal that can be detected with a fast detector. The comparison of the images of various cell lines at different cell-cycle stages obtained by OB polarization microscopy and fluorescence confocal images shows that an endogenous polarimetric contrast arizes on compacted chromosomes during cell division. This technique paves the way to label-free real-time polarization imaging of mitotic chromosomes with further potential applications in histology and cancer diagnosis.
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