Department of Epigenetics and Molecular Carcinogenesis, Center for Cancer Epigenetics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
Sci Rep. 2017 Jun 19;7(1):3779. doi: 10.1038/s41598-017-03610-2.
Mass cytometry presents an exceptional opportunity to interrogate the biology of highly heterogeneous cell populations, owing to the ability to collect highly parametric proteomic data at a single cell level. However, sample-to-sample variability, due to antibody staining and/or instrument sensitivity, can introduce substantial artifacts into the data, which can in turn lead to erroneous conclusions. This variability can be eliminated by sample barcoding which enables samples to be pooled, stained and run simultaneously. Existing mass cytometry barcoding approaches require time intensive labeling, reduce the number of biologically meaningful parameters and/or rely on expensive reagents. We present an approach utilizing monoisotopic cisplatin to perform cell barcoding that does not require cell permeabilization, can be completed in 10 minutes and can be utilized in combination with existing barcoding techniques to greatly increase the number of samples which can be multiplexed to improve throughput and consistency.
质谱流式细胞术能够在单细胞水平上获取高度参数化的蛋白质组学数据，为高度异质细胞群体的生物学研究提供了绝佳的机会。然而，由于抗体染色和/或仪器灵敏度的不同，样本间的变异性会给数据带来大量伪影，进而导致错误的结论。通过样本条形码化，可以消除这种变异性，从而实现样本的混合、染色和同时运行。现有的质谱流式细胞术条形码技术需要耗时的标记，减少了有生物学意义的参数数量，或者依赖昂贵的试剂。我们提出了一种利用顺铂的同位素标记来进行细胞条形码化的方法，该方法不需要细胞通透化，可以在 10 分钟内完成，并且可以与现有的条形码技术结合使用，从而大大增加可以进行多重化以提高通量和一致性的样本数量。
