Free-space angular-chirp-enhanced delay (FACED) is an ultrafast laser-scanning technique that allows for high imaging speed at the scale orders of magnitude greater than the current technologies. However, this speed advantage has only been restricted to bright-field and fluorescence imaging—limiting the variety of image contents and hindering its applicability in image-based bioassay, which increasingly demands rich phenotypic readout at a large scale. Here, we present a new high-speed quantitative phase imaging (QPI) based on time-interleaved phase-gradient FACED image detection. We further integrate this system with a microfluidic flow cytometer platform that enables synchronized and co-registered single-cell QPI and fluorescence imaging at an imaging throughput of 77 000 cells/s with sub-cellular resolution. Combined with deep learning, this platform empowers comprehensive image-based profiling of single-cell biophysical phenotypes that can offer not only sufficient label-free power for cell-type classification but also cell-cycle phase tracking with high accuracy comparable to the gold-standard fluorescence method. This platform further enables correlative, compartment-specific single-cell analysis of the spatially resolved biophysical profiles at the throughput inaccessible with existing QPI methods. The high imaging throughput and content given by this multimodal FACED imaging system could open new opportunities in image-based single-cell analysis, especially systematic analysis that correlates the biophysical and biochemical information of cells, and provide new mechanistic insights into biophysical heterogeneities in many biological processes.

中文翻译：

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用于大规模单细胞形态学分析的多模态 FACED 成像

自由空间角啁啾增强延迟 (FACED) 是一种超快激光扫描技术，可实现比当前技术大几个数量级的高成像速度。然而，这种速度优势仅限于明场和荧光成像——限制了图像内容的多样性并阻碍了其在基于图像的生物测定中的适用性，这越来越需要大规模的丰富表型读数。在这里，我们提出了一种新的基于时间交错相位梯度 FACED 图像检测的高速定量相位成像 (QPI)。我们进一步将该系统与微流体流式细胞仪平台集成，该平台能够以 77 000 个细胞/秒的成像通量和亚细胞分辨率实现同步和共同注册的单细胞 QPI 和荧光成像。结合深度学习，该平台支持对单细胞生物物理表型进行全面的基于图像的分析，不仅可以为细胞类型分类提供足够的无标记能力，还可以提供与金标准荧光方法相媲美的高精度细胞周期相位跟踪。该平台进一步支持以现有 QPI 方法无法达到的吞吐量对空间分辨生物物理剖面进行相关的、特定于隔室的单细胞分析。这种多模态 FACED 成像系统提供的高成像通量和内容可以为基于图像的单细胞分析开辟新的机会，尤其是将细胞的生物物理和生化信息相关联的系统分析，并为许多生物学中的生物物理异质性提供新的机制见解。过程。