Catena Raúl, Özcan Alaz, Zivanovic Nevena, Bodenmiller Bernd
Institute of Molecular Life Sciences (IMLS), University of Zürich, Zürich, Switzerland.
Department of Biology, Biology Master's Program, ETH, Zürich, Switzerland.
Cytometry A. 2016 May;89(5):491-7. doi: 10.1002/cyto.a.22848. Epub 2016 Mar 28.
Mass cytometry facilitates high-dimensional, quantitative, single-cell analysis. The method for sample multiplexing in mass cytometry, called mass-tag cellular barcoding (MCB), relies on the covalent reaction of bifunctional metal chelators with intracellular proteins. Here, we describe the use of osmium and ruthenium tetroxides (OsO4 and RuO4 ) that bind covalently with fatty acids in the cellular membranes and aromatic amino acids in proteins. Both OsO4 and RuO4 rapidly reacted and allowed for MCB with live cells, crosslinked cells, and permeabilized cells. Given the covalent nature of the labeling reaction, isotope leaching was not observed. OsO4 and RuO4 were used in a 20-sample barcoding protocol together with palladium isotopes. As mass channels occupied by osmium and ruthenium are not used for antibody detection the number of masses effectively utilized in a single experiment is expanded. OsO4 and RuO4 can therefore be used as MCB reagents for a wide range of mass cytometry workflows. © 2016 International Society for Advancement of Cytometry.
质谱流式细胞术有助于进行高维、定量的单细胞分析。质谱流式细胞术中用于样本多重分析的方法，称为质量标签细胞条形码（MCB），它依赖于双功能金属螯合剂与细胞内蛋白质的共价反应。在此，我们描述了使用四氧化锇和四氧化钌（OsO4和RuO4），它们可与细胞膜中的脂肪酸以及蛋白质中的芳香族氨基酸发生共价结合。OsO4和RuO4都能迅速反应，并可用于对活细胞、交联细胞和通透细胞进行MCB分析。鉴于标记反应的共价性质，未观察到同位素浸出现象。OsO4和RuO4与钯同位素一起用于一个20样本的条形码方案中。由于锇和钌占据的质量通道不用于抗体检测，因此单次实验中有效利用的质量数得以增加。因此，OsO4和RuO4可用作多种质谱流式细胞术工作流程的MCB试剂。© 2016国际细胞计量学促进协会。
