Schulz Axel Ronald, Mei Henrik E
Mass Cytometry Lab, German Rheumatism Research Center (DRFZ), A Leibniz Institute, Berlin, Germany.
Methods Mol Biol. 2019;1989:93-108. doi: 10.1007/978-1-4939-9454-0_7.
Sample barcoding is a powerful method for harmonizing mass cytometry data. By assigning a unique combination of barcode labels to each cell sample, a set of individual samples can be pooled and further processed and acquired as a large, single sample. For assays that require uncompromised profiling of cell-surface markers on live cells, barcoding by metal-labeled antibodies targeting cell-surface epitopes is the barcoding approach of choice. Here we provide an optimized and validated protocol for cell-surface barcoding of ten PBMC samples with palladium-labeled β2-microglobulin (B2M) antibodies used in a 5-choose-2 barcoding scheme, for subsequent immune phenotyping by mass cytometry. We further provide details on the generation of palladium-labeled antibodies utilizing amine-reactive isothiocyanobenzyl-EDTA (ITCB-EDTA) that permits the implementation of antibody-based barcoding not interfering with lanthanide channels typically used for analyte detection in mass cytometry assays.
样本条形码技术是一种用于整合质谱流式细胞术数据的强大方法。通过为每个细胞样本分配条形码标签的独特组合，可以将一组单独的样本汇集起来，并作为一个大的单个样本进行进一步处理和采集。对于需要对活细胞表面标志物进行完整分析的检测，通过靶向细胞表面表位的金属标记抗体进行条形码编码是首选的条形码方法。在这里，我们提供了一种优化且经过验证的方案，用于使用钯标记的β2-微球蛋白（B2M）抗体对十个外周血单核细胞（PBMC）样本进行细胞表面条形码编码，该方案采用5选2条形码方案，随后通过质谱流式细胞术进行免疫表型分析。我们还进一步详细介绍了利用胺反应性异硫氰酸苄基-EDTA（ITCB-EDTA）生成钯标记抗体的方法，该方法允许实施基于抗体的条形码编码，而不会干扰质谱流式细胞术检测中通常用于分析物检测的镧系元素通道。
