Ciliary movements within the human airway are essential for maintaining a clean lung environment. Motile cilia have a characteristic ciliary beat frequency (CBF). However, CBF measurement with current video microscopic techniques can be error-prone due to the use of the single-point Fourier transformation, which is often biased for ciliary measurements. Herein, we describe a new video microscopy technique that harnesses a metric of motion-contrast imaging and image correlation for CBF analysis. It can provide objective and selective CBF measurements for individual motile cilia and generate CBF maps for the imaged area. The measurement performance of our methodology was validated with in vitro human airway organoid models that simulated an actual human airway epithelium. The CBF determined for the region of interest (ROI) was equal to that obtained with manual counting. The signal redundancy problem of conventional methods was not observed. Moreover, the obtained CBF measurements were robust to optical focal shifts, and exhibited spatial heterogeneity and temperature dependence. This technique can be used to evaluate ciliary movement in respiratory tracts and determine whether it is non-synchronous or aperiodic in patients. Therefore, our observations suggest that the proposed method can be clinically adapted as a screening tool to diagnose ciliopathies.

中文翻译：

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基于图像相关的方法评估人体气道类器官纤毛跳动频率


人体气道内的纤毛运动对于维持清洁的肺部环境至关重要。运动纤毛具有特征性纤毛跳动频率（CBF）。然而，由于使用单点傅里叶变换，当前视频显微技术的 CBF 测量可能容易出错，单点傅里叶变换通常对纤毛测量有偏差。在此，我们描述了一种新的视频显微镜技术，该技术利用运动对比成像和图像相关性的度量进行 CBF 分析。它可以为单个运动纤毛提供客观和选择性的 CBF 测量，并生成成像区域的 CBF 图。我们的方法的测量性能通过模拟实际人类气道上皮的体外人类气道类器官模型进行了验证。感兴趣区域 (ROI) 确定的 CBF 等于手动计数获得的 CBF。没有观察到传统方法的信号冗余问题。此外，获得的 CBF 测量对光学焦移具有鲁棒性，并且表现出空间异质性和温度依赖性。该技术可用于评估呼吸道中的纤毛运动，并确定患者的纤毛运动是否是不同步的或非周期性的。因此，我们的观察表明，所提出的方法可以在临床上用作诊断纤毛病的筛查工具。