Stabilizing Antibody Cocktails for Mass Cytometry
Stabilizing Antibody Cocktails for Mass Cytometry
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DOI：
10.1002/cyto.a.23781
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发表时间：
2019-08-01
期刊：
影响因子：
3.7
通讯作者：
Mei, Henrik E.
中科院分区：
文献类型：
--
作者：
Schulz, Axel R.;Baumgart, Sabine;Mei, Henrik E.
质谱流式细胞术越来越多地应用于大型免疫分析研究，涉及几天和多个地点的数据采集。为了通过计算分析从相应数据获得最大信息，已经开发了几种技术来最小化大量细胞计数数据集中的噪声，例如样品库、标准化仪器设置、样品条形码化和信号归一化。然而，由同位素标记的抗体组成的混合物的重复制备仍然是一个显着的错误来源。我们在这里表明，预混合的抗体混合物不能提供预期的染色模式时，在4摄氏度储存4周。作为一种解决方案，我们开发并测试了一种用于质量细胞术的高度多重抗体混合物的冷冻保存方法，包括镧系元素、钯和铂缀合物，当在低于-80 ℃的温度下储存时，其产生稳定的染色模式至少9个月。使用抗体混合物的冷冻等分试样是一种经济且灵活的方法，可显著提高大规模细胞计数研究中的数据一致性，其中重复染色/测量周期跨越数天或涉及多个数据采集站点。(C)2019年国际细胞计数促进学会
Mass cytometry is increasingly employed in larger immune profiling studies involving data acquisitions across several days and multiple sites. For gaining a maximum of information from respective data by computational analyses, several techniques have been developed to minimize noise in mass cytometric data sets, such as sample banking, standardized instrument setup, sample barcoding, and signal normalization. However, the repeated preparation of cocktails composed of isotope-tagged antibodies remained a significant source of error.We here show that premixed antibody cocktails fail to deliver expected staining patterns when stored at 4 degrees C for 4 weeks. As a solution, we developed and tested a cryopreservation method for highly multiplexed antibody cocktails for mass cytometry including lanthanide, palladium, and platinum conjugates that yielded stable staining patterns for at least 9 months when stored at temperatures below -80 degrees C. Using frozen aliquots of antibody cocktails is an economic and flexible approach to significantly improve data consistency in large mass cytometry studies with repetitive staining/measurement cycles spanning several days or involving multiple data acquisition sites. (C) 2019 International Society for Advancement of Cytometry