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Optical coherence microscopy as a novel non-invasive method for the 4D live imaging of early mammalian embryos

机译:光学相干显微镜是一种新颖的非侵入性的早期哺乳动物胚胎4D实时成像方法

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摘要

Imaging of living cells based on traditional fluorescence and confocal laser scanning microscopy has delivered an enormous amount of information critical for understanding biological processes in single cells. However, the requirement for a high numerical aperture and fluorescent markers still limits researchers’ ability to visualize the cellular architecture without causing short- and long-term photodamage. Optical coherence microscopy (OCM) is a promising alternative that circumvents the technical limitations of fluorescence imaging techniques and provides unique access to fundamental aspects of early embryonic development, without the requirement for sample pre-processing or labeling. In the present paper, we utilized the internal motion of cytoplasm, as well as custom scanning and signal processing protocols, to effectively reduce the speckle noise typical for standard OCM and enable high-resolution intracellular time-lapse imaging. To test our imaging system we used mouse and pig oocytes and embryos and visualized them through fertilization and the first embryonic division, as well as at selected stages of oogenesis and preimplantation development. Because all morphological and morphokinetic properties recorded by OCM are believed to be biomarkers of oocyte/embryo quality, OCM may represent a new chapter in imaging-based preimplantation embryo diagnostics.
机译:基于传统的荧光和共聚焦激光扫描显微镜对活细胞进行成像,已提供了大量信息,这些信息对于理解单个细胞的生物学过程至关重要。但是,对高​​数值孔径和荧光标记的要求仍然限制了研究人员可视化细胞结构而不会造成短期和长期光损伤的能力。光学相干显微镜(OCM)是一种有前途的替代方案,它可以规避荧光成像技术的技术局限性,并提供对早期胚胎发育基本方面的独特访问,而无需进行样品预处理或标记。在本文中,我们利用细胞质的内部运动以及自定义的扫描和信号处理协议来有效降低标准OCM的典型斑点噪声,并实现高分辨率的细胞内延时成像。为了测试我们的成像系统,我们使用了小鼠和猪的卵母细胞和胚胎,并通过受精和第一个胚胎分裂以及在卵子发生和植入前发育的选定阶段对其进行了可视化。由于OCM记录的所有形态和形态动力学特性均被认为是卵母细胞/胚胎质量的生物标记,因此OCM可能代表了基于成像的植入前胚胎诊断的新篇章。

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