A potential hurdle when applying mass cytometry to the field study setting is the streamlining of sample collection while at the same time protecting the integrity of important cell epitopes. Whole blood preservation kits applying fixation and/or permeabilization agents are increasingly used in clinical trials to preserve leukocytes needed for downstream analysis. We here present a structured overview of leukocyte surface marker detectability in samples processed with four commercially available whole blood preservation kits; 1) Proteomic Stabilizer, 2) Stable-Lyse V2 and Stable-Store V2, 3) Cytodelics and 4) Lyse/Fix Buffer, as well as in samples treated with buffers included in Mass-tag Cellular Barcoding kits. Isolated leukocytes were stained with a 28-marker panel (including 7 chemokine receptors) of metal-conjugated antibodies and analysed on a mass cytometer. Exploration of the data by manual gating and viSNE analysis showed that although many markers were similarly detected across all sample conditions, most of the chemokine receptors in our panel, particularly CXCR3, CCR4, CCR6 and CXCR5, were incorrectly detected in the preserved samples and thus incompatible with the fixation and permeabilization agents found in whole blood preservation kits and in buffers used prior to barcoding.

中文翻译：

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全血保存方法会改变细胞计数实验中趋化因子受体的检测。

在将大规模细胞计数法应用于现场研究环境时，潜在的障碍是精简样品收集，同时保护重要细胞表位的完整性。应用固定剂和/或通透剂的全血保存试剂盒在临床试验中越来越多地用于保存下游分析所需的白细胞。我们在此提供了使用四种市售全血保存试剂盒处理的样品中白细胞表面标志物可检测性的结构化概述；1）蛋白质组稳定剂，2）稳定液V2和稳定储存V2、3）细胞保质液和4）裂解/固定缓冲液，以及用Mass-tag Cellular Barcoding试剂盒中包含的缓冲液处理的样品。分离的白细胞用结合有金属的抗体的28个标记组（包括7个趋化因子受体）染色，并在质谱仪上进行分析。通过手动门控和viSNE分析对数据进行的探索表明，尽管在所有样品条件下都检测到许多标记，但我们小组中的大多数趋化因子受体，特别是CXCR3，CCR4，CCR6和CXCR5，在保存的样品中均被错误地检测到，因此与全血保鲜盒和条形码扫描前使用的缓冲液中的固定剂和透化剂不兼容。