High-throughput single cell motility analysis using nanowell-in-microwells

IF 5.4 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2025-07-04 DOI:10.1039/D5LC00478K
Pan Deng, Wenze Lyu, Deasung Jang, Kerryn Matthews, Simon P. Duffy and Hongshen Ma
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引用次数: 0

Abstract

Cell motility is important to many biological processes including cancer, immune response, and tissue repair. Conventional assays measure bulk cell motility, potentially overlooking important heterogeneity and missing important high motility subpopulations. Here, we introduce a high-throughput single-cell motility assay using nanowell-in-microwell plates to precisely track single cell position and analyze their migratory trajectories. By physically confining individual cells in nanowells, we eliminate cell–cell interactions and simplify cell segmentation and tracking. Using this platform, we characterized the motility of single cells across different culture conditions to identify distinct motility phenotypes. Single-cell trajectory analysis revealed pronounced directional persistence, with cells predominantly maintaining their direction of travel and trajectory along nanowell boundaries. Additionally, our approach facilitates the generation of labeled image datasets suitable for AI models to rapidly identify cell motility phenotypes from single-cell images. Together, our platform provides a robust, scalable method to analyze cell motility phenotypes and migration behavior at single-cell resolution.

Abstract Image

微孔中纳米孔高通量单细胞运动分析
细胞运动对许多生物过程都很重要,包括癌症、免疫反应和组织修复。传统的测定方法测量大细胞运动性,潜在地忽略了重要的异质性和缺失了重要的高运动性亚群。在这里,我们介绍了一种高通量单细胞运动测定,使用微孔板中的纳米孔来精确跟踪单细胞的位置并分析它们的迁移轨迹。通过物理上将单个细胞限制在纳米孔中,我们消除了细胞间的相互作用,简化了细胞分割和跟踪。利用这个平台,我们表征了单细胞在不同培养条件下的运动性,以确定不同的运动性表型。单细胞轨迹分析显示了明显的定向持久性,细胞主要沿着纳米井边界保持其行进方向和轨迹。此外,我们的方法有助于生成适合AI模型从单细胞图像中分类细胞运动表型的标记图像数据集。总之,我们的平台提供了一个强大的,可扩展的方法来分析单细胞分辨率的细胞运动表型和迁移行为。
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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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