All-digital histopathology by infrared-optical hybrid microscopy
All-digital histopathology by infrared-optical hybrid microscopy
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DOI：
10.1073/pnas.1912400117
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发表时间：
2020-02-18
影响因子：
11.1
通讯作者：
Bhargava, Rohit
中科院分区：
文献类型：
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作者：
Schnell, Martin;Mittal, Shachi;Bhargava, Rohit
生物医学样品的光学显微镜需要染色方面的专业知识以可视化结构和成分。中红外 (mid-IR) 光谱成像通过探测分子成分的基本振动模式提供无标记分子记录和虚拟染色。这种定量信号可以与机器学习相结合，使显微镜能够应用于从癌症诊断到法医学的各个领域。然而，普通光学成像组件对红外光的吸收使得中红外光与现代光学显微镜以及几乎所有生物医学研究和临床工作流程不兼容。在这里，我们概念化了一种红外光学混合 (IR-OH) 方法，该方法基于光学显微镜灵敏地测量分子组成，并可对吸收引起的样品膨胀进行宽视场干涉检测。我们证明 IR-OH 在覆盖范围（10 倍）、空间分辨率（四倍）和光谱一致性（通过减轻散射影响）方面超过了最先进的红外显微镜。这些进步的综合影响使得未染色的乳腺组织切片的完整幻灯片红外吸收图像可以在可见显微镜平台上进行。我们进一步表明，自动组织病理学分割和计算染色（不锈钢）图像的生成是可能的，解决了与当前病理学协议相当的颜色和空间细节的形态特征，但没有染色或人类解释。 IR-OH 与临床和研究病理学实践兼容，可以成为不锈钢全数字病理学传统基于染色方案的经济高效的替代方案。
Optical microscopy for biomedical samples requires expertise in staining to visualize structure and composition. Midinfrared (mid-IR) spectroscopic imaging offers label-free molecular recording and virtual staining by probing fundamental vibrational modes of molecular components. This quantitative signal can be combined with machine learning to enable microscopy in diverse fields from cancer diagnoses to forensics. However, absorption of IR light by common optical imaging components makes mid-IR light incompatible with modern optical microscopy and almost all biomedical research and clinical workflows. Here we conceptualize an IR-optical hybrid (IR-OH) approach that sensitively measures molecular composition based on an optical microscope with wide-field interferometric detection of absorption-induced sample expansion. We demonstrate that IR-OH exceeds state-of-the-art IR microscopy in coverage (10-fold), spatial resolution (fourfold), and spectral consistency (by mitigating the effects of scattering). The combined impact of these advances allows full slide infrared absorption images of unstained breast tissue sections on a visible microscope platform. We further show that automated histopathologic segmentation and generation of computationally stained (stainless) images is possible, resolving morphological features in both color and spatial detail comparable to current pathology protocols but without stains or human interpretation. IR-OH is compatible with clinical and research pathology practice and could make for a cost-effective alternative to conventional stain-based protocols for stainless, all-digital pathology.