使用单细胞细胞毒性试验评估免疫靶细胞相互作用的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-10 DOI:10.1016/j.xpro.2024.103558

Zhihao Wei, Konglan Lin, Min Huang, Shicheng Su, Yiwen Lu

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引用次数: 0

摘要

基于流式细胞术的标准检测方法可以确定免疫效应细胞的细胞毒性，但对细胞毒性的动态过程进行监测具有挑战性。在这里，我们提出了一种使用自动显微镜用微孔阵列连续观察自然杀伤（NK）细胞介导的细胞毒性的方案。我们描述了分离和标记原代NK细胞，将细胞装载到微孔阵列，监测目标孔和图像分析的步骤。该方案有利于在单细胞水平上观察免疫靶细胞相互作用的动力学。有关该协议的使用和执行的完整细节，请参阅Li等人1。

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Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay.

Standard flow cytometry-based assays can determine the cytotoxicity of immune effector cells, but it is challenging to monitor the dynamic processes of cytotoxicity. Here, we present a protocol for continuous observation of natural killer (NK) cell-mediated cytotoxicity with microwell arrays using an automated microscope. We describe steps for isolating and labeling primary NK cells, loading cells onto microwell arrays, monitoring target wells, and image analysis. This protocol facilitates observation of the dynamics of immune-target cell interactions at the single-cell level. For complete details on the use and execution of this protocol, please refer to Li et al.¹.

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