标题： 数字失焦像差干涉用于自动光学显微镜 显示英文标题

标题： Digital defocus aberration interference for automated optical microscopy

摘要： 自动化在光学显微镜中对于在广泛生物医学应用中实现高通量成像至关重要。 在自动系统的基本组件中，鲁棒的自动对焦在保持单平面和体积成像的图像质量方面起着关键作用。 然而，传统的自动对焦方法通常在实现复杂性、跨样本类型的泛化能力有限、与厚标本不兼容以及反馈速度慢方面存在困难。 我们最近发现了一种现象，即从两个角度照明获取的两幅图像的数字求和傅里叶谱在样品失焦时表现出类似干涉的条纹调制。 这些数字条纹通过基于物理的关系直接与失焦相关。 基于这一原理，我们开发了一种自动、高效且泛化的失焦检测方法，称为数字失焦像差干涉（DAbI）。 采用简单的双LED设置，DAbI可以量化薄样品的失焦距离范围为212倍的焦深（DoF），厚标本为300倍。 当与复场成像结合时，它还可以将成像系统的自然焦深扩展20倍。 我们在明场、复场、折射率、共聚焦和宽场荧光成像中展示了DAbI的多功能应用，确立了其作为自动化高通量光学显微镜有前景的解决方案。 显示英文摘要

摘要： Automation in optical microscopy is critical for enabling high-throughput imaging across a wide range of biomedical applications. Among the essential components of automated systems, robust autofocusing plays a pivotal role in maintaining image quality for both single-plane and volumetric imaging. However, conventional autofocusing methods often struggle with implementation complexity, limited generalizability across sample types, incompatibility with thick specimens, and slow feedback. We recently discovered a phenomenon that the digitally summed Fourier spectrum of two images acquired from two-angle illumination exhibits interference-like fringe modulation when the sample is defocused. These digital fringes correlate directly with defocus through a physics-based relation. Based on this principle, we developed an automatic, efficient, and generalizable defocus detection method termed digital defocus aberration interference (DAbI). Implemented with a simple two-LED setup, DAbI can quantify the defocus distance over a range of 212 times the depth-of-field (DoF) for thin samples and 300 times for thick specimens. It can additionally extend the natural DoF of the imaging system by 20 folds when integrated with complex-field imaging. We demonstrated the versatile applications of DAbI on brightfield, complex-field, refractive index, confocal, and widefield fluorescence imaging, establishing it as a promising solution for automated, high-throughput optical microscopy.