可视化质谱流式细胞术信号背景以实现最佳核心面板定制和信号阈值门控。

Visualization of Mass Cytometry Signal Background to Enable Optimal Core Panel Customization and Signal Threshold Gating.

作者信息

Au-Yeung Amelia, Takahashi Chikara, Mathews W Rodney, O'Gorman William E

机构信息

OMNI Biomarkers, Development Sciences, Genentech, South San Francisco, CA, USA.

出版信息

Methods Mol Biol. 2019;1989:35-45. doi: 10.1007/978-1-4939-9454-0_3.

DOI:10.1007/978-1-4939-9454-0_3

PMID:31077097

Abstract

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.

摘要

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。