The observation of biological structures in live cells beyond the diffraction limit with super-resolution fluorescence microscopy is limited by the ability of fluorescence probes to permeate live cells and the effect of these probes, which are often toxic, on cellular behavior. Here we present a coherent confocal light scattering and absorption spectroscopic microscopy that for the first time enables the use of large numerical aperture optics to characterize structures in live cells down to 10 nm spatial scales, well beyond the diffraction limit. Not only does this new capability allow high-resolution microscopy with light scattering contrast, but it can also be used with almost any light scattering spectroscopic application that employs lenses. We demonstrate that the coherent light scattering contrast based technique allows continuous temporal tracking of the transition from noncancerous to an early cancerous state in live cells, without exogenous markers. We also use the technique to sense differences in the aggressiveness of cancer in live cells and for label-free identification of different grades of cancer in resected tumor tissues.

中文翻译：

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相干共焦光散射光谱显微镜评估活细胞和组织中的癌症进展和侵袭性

用超分辨率荧光显微镜观察活细胞中超出衍射极限的生物结构受到荧光探针渗透活细胞的能力以及这些探针（通常是有毒的）对细胞行为的影响的限制。在这里，我们提出了一种相干共焦光散射和吸收光谱显微镜，它首次能够使用大数值孔径光学来表征低至 10 nm 空间尺度的活细胞结构，远远超出衍射极限。这种新功能不仅允许具有光散射对比度的高分辨率显微镜，而且它还可以用于几乎任何使用透镜的光散射光谱应用。我们证明了基于相干光散射对比的技术允许在没有外源标记的情况下连续时间跟踪活细胞中从非癌性状态到早期癌性状态的转变。我们还使用该技术来感知活细胞中癌症侵袭性的差异，并在切除的肿瘤组织中对不同级别的癌症进行无标记鉴定。