24-color (30-marker) antibody panel for deep immunophenotyping of immune cells in human peripheral blood

This OMIP provides a detailed description of a 24-color (30-markers) full spectrum flow cytometry panel to identify all major immune compartments of blood, neutrophils, basophils, eosinophils as well as T, B, MAIT, Treg, NK, NKT-like, ?? T, monocytes and dendritic cells, basophils, ILCs and their associated subsets. Activation, differentiation, check point and senescence markers were employed to provide an in-depth immune profiling in a single sample.
Conclusion:
Full Spectrum Flow Cytometry allows detection of 30 markers simultaneously with good population resolution, allowing comprehensive in-depth immune cell profiling within a single biological sample.
Notes:
Unmixed data file with FlowJo workspace datafile used in OMIP as an example. Daily QC was performed using SpectroFlo® QC beads prior to acquiring samples to ensure that the cytometer was performing optimally. Daily QC assessed the instrument’s optical alignment and the system performance drift by measuring %rCVs and gains needed to place the beads at the target locations established for each detector. During the typical QC protocol, Laser delays and area scaling factors were also optimized, and gain settings adjusted to account for day-to-day instrument variability.

本开放免疫表型分型面板（Open Immunophenotyping Panel，OMIP）详细描述了一套24色（30标记）全光谱流式细胞术检测面板，可用于鉴定血液中所有主要免疫组分：中性粒细胞、嗜碱性粒细胞、嗜酸性粒细胞，以及T细胞、B细胞、黏膜相关恒定T细胞（MAIT）、调节性T细胞（Treg）、自然杀伤细胞（NK）、类自然杀伤T细胞（NKT-like）、?? T细胞、单核细胞、树突状细胞、嗜碱性粒细胞、固有淋巴细胞（ILCs）及其相关亚群。该面板采用活化、分化、免疫检查点及衰老相关标记物，可实现单样本的深度免疫谱分析。
结论：
全光谱流式细胞术可同时检测30个标记物，且具备优异的群体分辨能力，可在单份生物样本中实现全面深入的免疫细胞谱分析。
注意事项：
本OMIP以未拆分数据文件及配套的FlowJo工作区数据文件作为示例。在样本获取前，需使用SpectroFlo®质量控制（Quality Control，QC）微球开展日常质量控制，以确保流式细胞仪处于最佳运行状态。日常质量控制通过测量相对变异系数百分比（%rCVs）及将微球定位至各检测器预设目标位置所需的增益值，评估仪器的光学对准情况与系统性能漂移情况。在标准质量控制流程中，还需优化激光延迟参数与面积缩放因子，并调整增益设置以抵消仪器日常运行的性能波动。