This 40‐color flow cytometry‐based panel was developed for in‐depth immunophenotyping of the major cell subsets present in human peripheral blood. Sample availability can often be limited, especially in cases of clinical trial material, when multiple types of testing are required from a single sample or timepoint. Maximizing the amount of information that can be obtained from a single sample not only provides more in‐depth characterization of the immune system but also serves to address the issue of limited sample availability. The panel presented here identifies CD4 T cells, CD8 T cells, regulatory T cells, γδ T cells, NKT‐like cells, B cells, NK cells, monocytes and dendritic cells. For each specific cell type, the panel includes markers for further characterization by including a selection of activation and differentiation markers, as well as chemokine receptors. Moreover, the combination of multiple markers in one tube might lead to the discovery of new immune phenotypes and their relevance in certain diseases. Of note, this panel was designed to include only surface markers to avoid the need for fixation and permeabilization steps. The panel can be used for studies aimed at characterizing the immune response in the context of infectious or autoimmune diseases, monitoring cancer patients on immuno‐ or chemotherapy, and discovery of unique and targetable biomarkers. Different from all previously published OMIPs, this panel was developed using a full spectrum flow cytometer, a technology that has allowed the effective use of 40 fluorescent markers in a single panel. The panel was developed using cryopreserved human peripheral blood mononuclear cells (PBMC) from healthy adults (Table 1). Although we have not tested the panel on fresh PBMCs or whole blood, it is anticipated that the panel could be used in those sample preparations without further optimization. @ 2020 Cytek Biosciences, Inc. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

中文翻译：

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OMIP-069：用于人外周血中主要细胞亚群深度免疫表型分析的 40 色全谱流式细胞术面板。

这种基于 40 色流式细胞术的面板是为深入分析人类外周血中存在的主要细胞亚群而开发的。样本可用性通常是有限的，特别是在临床试验材料的情况下，当需要从单个样本或时间点进行多种类型的测试时。最大化从单个样本中获得的信息量不仅可以更深入地表征免疫系统，还可以解决样本可用性有限的问题。这里展示的面板识别了 CD4 T 细胞、CD8 T 细胞、调节性 T 细胞、γδ T 细胞、NKT 样细胞、B 细胞、NK 细胞、单核细胞和树突状细胞。对于每种特定的细胞类型，该面板包括用于进一步表征的标记，包括一系列激活和分化标记，以及趋化因子受体。此外，在一个管中组合多个标记可能会导致发现新的免疫表型及其与某些疾病的相关性。值得注意的是，该面板旨在仅包含表面标记，以避免需要固定和通透步骤。该面板可用于旨在表征感染或自身免疫疾病背景下的免疫反应、监测接受免疫或化疗的癌症患者以及发现独特且可靶向的生物标志物的研究。与之前发布的所有 OMIP 不同，该面板是使用全光谱流式细胞仪开发的，该技术允许在单个面板中有效使用 40 种荧光标记。该面板是使用来自健康成人的冷冻保存的人外周血单核细胞 (PBMC) 开发的（表 1）。尽管我们尚未在新鲜 PBMC 或全血上测试该面板，但预计该面板无需进一步优化即可用于这些样品制备。@ 2020 Cytek Biosciences, Inc.细胞学 A 部分由 Wiley Periodicals LLC 代表国际细胞学促进会出版。