Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay
Summary: 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 array...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007238 |
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| _version_ | 1841545877271871488 |
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| author | Zhihao Wei Konglan Lin Min Huang Shicheng Su Yiwen Lu |
| author_facet | Zhihao Wei Konglan Lin Min Huang Shicheng Su Yiwen Lu |
| author_sort | Zhihao Wei |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-06f439674add44e89f6fb86dcd74f91c |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-06f439674add44e89f6fb86dcd74f91c2025-01-11T06:41:59ZengElsevierSTAR Protocols2666-16672025-03-0161103558Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assayZhihao Wei0Konglan Lin1Min Huang2Shicheng Su3Yiwen Lu4Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, ChinaGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, ChinaGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, ChinaGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Department of Immunology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510080, ChinaGuangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007238Cell-based AssaysCancerClinical ProtocolImmunology |
| spellingShingle | Zhihao Wei Konglan Lin Min Huang Shicheng Su Yiwen Lu Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay STAR Protocols Cell-based Assays Cancer Clinical Protocol Immunology |
| title | Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay |
| title_full | Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay |
| title_fullStr | Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay |
| title_full_unstemmed | Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay |
| title_short | Protocol for assessing immune-target cell interactions using a single-cell cytotoxicity assay |
| title_sort | protocol for assessing immune target cell interactions using a single cell cytotoxicity assay |
| topic | Cell-based Assays Cancer Clinical Protocol Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007238 |
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