Palladium-based mass tag cell barcoding with a doublet-filtering scheme and single-cell deconvolution algorithm.
Palladium-based mass tag cell barcoding with a doublet-filtering scheme and single-cell deconvolution algorithm.
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DOI：
10.1038/nprot.2015.020
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发表时间：
2015-02
期刊：
影响因子：
14.8
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质量标签细胞条形码 (MCB) 用独特的组合条形码标记单个细胞样本，然后将它们汇集起来作为单个多重样本进行处理和测量。 MCB方法消除了样品之间抗体染色和仪器灵敏度的差异，减少了抗体消耗，并缩短了仪器测量时间。在这里，我们提出了一种优化的 MCB 协议，与之前描述的方法相比有一些改进。使用基于钯的标记试剂扩大了可用于质谱流式分析的测量通道的数量，并减少了对基于镧系元素的抗体测量的干扰。错误检测组合条形码方案允许识别细胞双联体并将其从分析中删除。基于单细胞而不是基于群体的去条形码算法提高了样本去卷积的准确性和效率。该去条形码算法已被打包到软件中，可实现快速且无偏差的样本去卷积。 MCB 程序需要 3-4 小时，不包括每百万细胞约 1 小时的样本采集时间。
Mass-tag cell barcoding (MCB) labels individual cell samples with unique combinatorial barcodes, after which they are pooled for processing and measurement as a single multiplexed sample. The MCB method eliminates variability between samples in antibody staining and instrument sensitivity, reduces antibody consumption, and shortens instrument measurement time. Here, we present an optimized MCB protocol with several improvements over previously described methods. The use of palladium-based labeling reagents expands the number of measurement channels available for mass cytometry and reduces interference with lanthanide-based antibody measurement. An error-detecting combinatorial barcoding scheme allows cell doublets to be identified and removed from the analysis. A debarcoding algorithm that is single cell-based rather than population-based improves the accuracy and efficiency of sample deconvolution. This debarcoding algorithm has been packaged into software that allows rapid and unbiased sample deconvolution. The MCB procedure takes 3–4 h, not including sample acquisition time of ~1 h per million cells.