Functional phenotyping of hepatic lymphocytes in murine MASH by mass cytometry.

Functional phenotyping of hepatic lymphocytes in murine MASH by mass cytometry.
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小鼠MASH中肝淋巴细胞功能表型的质谱细胞术分析。

DOI:
10.1016/j.xpro.2023.102743
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发表时间:
2023-12-15
期刊:
影响因子:
--
通讯作者:
Revelo, Xavier S.
Revelo, Xavier S.
中科院分区:
其他
文献类型:
--
作者:
Barrow, Fanta;Fredrickson, Gavin;Wang, Haiguang;Revelo, Xavier S.

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由B和T淋巴细胞等免疫细胞驱动的肝脏炎症是代谢功能障碍相关性脂肪性肝炎(MASH)的一个显著特征。在这里,我们详细介绍了一种通过飞行时间(CyTOF)程序对MASH小鼠肝淋巴细胞表型进行检测的强大的细胞学方法。我们采用了抗体的定制金属偶联,肝脏淋巴细胞的分离,细胞表面和细胞内的染色,以及数据采集。该方案克服了传统流式细胞术的局限性,可容纳多达40个标记进行全面的免疫表型鉴定。有关该协议的使用和执行的完整细节,请参考Barrow等人。一份完整的肝脏免疫细胞细胞表型分析流程详细说明了结合金属标记抗体、细胞表面染色和细胞质/分泌蛋白样本采集的步骤,以及代表性结果的发布说明:实施任何实验方案都需要遵守当地实验室安全和伦理方面的机构指南。由B和T淋巴细胞等免疫细胞驱动的肝脏炎症是代谢功能障碍相关性脂肪性肝炎(MASH)的一个显著特征。在这里,我们详细介绍了一种通过飞行时间(CyTOF)程序对MASH小鼠肝淋巴细胞表型进行检测的强大的细胞学方法。我们采用了抗体的定制金属偶联,肝脏淋巴细胞的分离,细胞表面和细胞内的染色,以及数据采集。该方案克服了传统流式细胞术的局限性,可容纳多达40个标记进行全面的免疫表型鉴定。
Hepatic inflammation, driven by immune cells such as B and T lymphocytes, is a hallmark feature of metabolic dysfunction-associated steatohepatitis (MASH). Here, we detail a robust cytometry by time-of-flight (CyTOF) procedure to phenotype hepatic lymphocytes from mice with MASH. We employ custom metal conjugation of antibodies, isolation of hepatic lymphocytes, cell surface and intracellular staining, and data acquisition. This protocol overcomes the limitations of traditional flow cytometry by accommodating up to 40 markers for comprehensive immune phenotyping. For complete details on the use and execution of this protocol, please refer to Barrow et al. A complete workflow for the phenotyping of hepatic immune cells by CyTOF Detailed steps for the conjugation of metal-tagged antibodies Staining of cell surface and cytoplasmic/secreted proteins Sample acquisition on a mass cytometer and representative results Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Hepatic inflammation, driven by immune cells such as B and T lymphocytes, is a hallmark feature of metabolic dysfunction-associated steatohepatitis (MASH). Here, we detail a robust cytometry by time-of-flight (CyTOF) procedure to phenotype hepatic lymphocytes from mice with MASH. We employ custom metal conjugation of antibodies, isolation of hepatic lymphocytes, cell surface and intracellular staining, and data acquisition. This protocol overcomes the limitations of traditional flow cytometry by accommodating up to 40 markers for comprehensive immune phenotyping.
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