通过定量相位成像和低折射率微孔中的限制来跟踪谱系质量。
Tracking of lineage mass via quantitative phase imaging and confinement in low refractive index microwells.
发表日期：2024 Aug 27
作者：
Jingzhou Zhang, Justin Griffin, Koushik Roy, Alexander Hoffmann, Thomas A Zangle
来源：
LAB ON A CHIP
摘要：
细胞谱系的测量对于从发育生物学到癌症生物学等各种基本生物学问题至关重要。然而，准确的谱系追踪需要近乎完美的细胞追踪，由于成像过程中的细胞运动，这可能具有挑战性。在这里，我们展示了微加工、成像和图像处理方法的集成，以展示细胞谱系追踪的平台。我们使用定量相位成像（QPI），这是一种量化细胞质量的无标记成像方法。这给出了一个附加参数，细胞质量，可用于提高跟踪精度。我们将谱系限制在微孔内，以减少细胞粘附到由低折射率聚合物制成的侧壁。这也使得微孔本身可以作为 QPI 的参考，甚至可以在汇合的微孔中测量细胞质量。我们展示了这种方法在永生化贴壁和非贴壁细胞系以及离体培养的刺激原代 B 细胞中的应用。总体而言，我们的方法可以在可针对不同细胞类型进行定制的平台中实现谱系跟踪或谱系质量测量。
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 ex vivo. Overall, our approach enables lineage tracking, or measurement of lineage mass, in a platform that can be customized to varied cell types.