Department of Chemical Engineering, University of Utah, USA.
Division of Microbiology and Immunology, Department of Pathology, School of Medicine, University of Utah, USA.
Lab Chip. 2024 Sep 10;24(18):4440-4449. doi: 10.1039/d4lc00389f.
Measurements of cell lineages are central to a variety of fundamental biological questions, ranging from developmental to cancer biology. However, accurate lineage tracing requires nearly perfect cell tracking, which can be challenging due to cell motion during imaging. Here we demonstrate the integration of microfabrication, imaging, and image processing approaches to demonstrate a platform for cell lineage tracing. We use quantitative phase imaging (QPI), a label-free imaging approach that quantifies cell mass. This gives an additional parameter, cell mass, that can be used to improve tracking accuracy. We confine lineages within microwells fabricated to reduce cell adhesion to sidewalls made of a low refractive index polymer. This also allows the microwells themselves to serve as references for QPI, enabling measurement of cell mass even in confluent microwells. We demonstrate application of this approach to immortalized adherent and nonadherent cell lines as well as stimulated primary B cells cultured . Overall, our approach enables lineage tracking, or measurement of lineage mass, in a platform that can be customized to varied cell types.
细胞谱系的测量是各种基础生物学问题的核心，包括发育生物学和癌症生物学。然而，精确的谱系追踪需要近乎完美的细胞跟踪，这由于成像过程中的细胞运动而具有挑战性。在这里，我们展示了微加工、成像和图像处理方法的集成，以展示用于细胞谱系追踪的平台。我们使用定量相位成像（QPI），一种定量细胞质量的无标记成像方法。这提供了一个额外的参数，即细胞质量，可以用来提高跟踪精度。我们将谱系限制在微井内，这些微井是为了减少细胞与低折射率聚合物制成的侧壁的粘附而制造的。这也允许微井本身作为 QPI 的参考，即使在密集的微井中也可以测量细胞质量。我们展示了该方法在永生化贴壁和非贴壁细胞系以及刺激的原代 B 细胞培养物中的应用。总的来说，我们的方法可以在一个可以针对不同细胞类型进行定制的平台上进行谱系追踪或谱系质量测量。
