Au-Yeung Amelia, Takahashi Chikara, Mathews W Rodney, O'Gorman William E
OMNI Biomarkers, Development Sciences, Genentech, South San Francisco, CA, USA.
Methods Mol Biol. 2019;1989:35-45. doi: 10.1007/978-1-4939-9454-0_3.
Signal interference or overlap in mass cytometry is minimal compared to flow cytometry but must still be considered for optimal panel design and assay sensitivity. Here we describe a procedure for evaluating signal interference dynamics in the context of a 25-parameter core immunophenotyping panel. Specifically, a mass-minus-many (MMM) approach was used to assess background signals in "empty" or "blank" channels intended for further customization. Through this approach cell type-specific variability in signal background is revealed. Further panel customization can thus be performed with an understanding of cell type and channel-specific background levels to enable rational panel design and the objective delineation of gating thresholds during analysis.
与流式细胞术相比，质谱流式细胞术中的信号干扰或重叠极小，但为了实现最佳的抗体组合设计和检测灵敏度，仍必须予以考虑。在此，我们描述了一种在25参数核心免疫表型分析抗体组合的背景下评估信号干扰动态的方法。具体而言，采用了多减一（MMM）方法来评估用于进一步定制的“空”通道或“空白”通道中的背景信号。通过这种方法，可以揭示信号背景中细胞类型特异性的变异性。因此，在了解细胞类型和通道特异性背景水平的情况下，可以进一步进行抗体组合定制，以实现合理的抗体组合设计，并在分析过程中客观地划定设门阈值。
