Development of a Click-Chemistry Reagent Compatible with Mass Cytometry.
Development of a Click-Chemistry Reagent Compatible with Mass Cytometry.
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DOI：
10.1038/s41598-018-25000-y
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发表时间：
2018-04-27
影响因子：
4.6
通讯作者：
van Berlo JH
中科院分区：
文献类型：
--
作者：
Shaklee J;Srivastava K;Brown H;Arriaga EA;Pierre VC;van Berlo JH
质谱细胞仪的最新发展已经允许同时检测来自单个细胞的40个或更多个独特参数。虽然类似于基于荧光团检测的流式细胞术，但质谱细胞术的一个关键区别特征是在质谱细胞仪中通过质谱法检测镧系元素的原子质量。其上级质量分辨率导致信号重叠的缺乏，从而允许每个单细胞中的分子特征的多参数检测大于流式细胞术的多参数检测，流式细胞术限于20个参数。不幸的是，大多数质谱检测依赖于镧系元素标记的抗体，这是理想的检测蛋白质，但不是其他类型的分子特征。为了进一步扩大可通过质谱细胞术检测的分子特征库，我们开发了一种镧系元素螯合的含叠氮化物的探针，其允许点击化学介导的靶分子标记。在细胞周期S期DNA合成过程中掺入胸苷类似物5-乙炔基-2 ′-脱氧尿苷（EdU）后，我们证明了此处引入的标记有铽-159（159 Tb）的探针通过铜催化的叠氮化物-炔Huisgen环加成（点击化学）与Edu反应。因此，通过检测159 Tb，可以使用质谱细胞计数法测定单细胞中的DNA合成。本文介绍的方法显示出与用于通过质谱细胞术测量单细胞中DNA合成的其他方法相似的灵敏度（真阳性率），并且与单细胞中其他参数的平行基于抗体的检测兼容。由于其通用性，点击化学在质谱细胞术中的使用扩展了可以通过质谱细胞术监测的分子靶标的类型。
The recent development of mass cytometry has allowed simultaneous detection of 40 or more unique parameters from individual single cells. While similar to flow cytometry, which is based on detection of fluorophores, one key distinguishing feature of mass cytometry is the detection of atomic masses of lanthanides by mass spectrometry in a mass cytometer. Its superior mass resolution results in lack of signal overlap, thereby allowing multiparametric detection of molecular features in each single cell greater than that of flow cytometry, which is limited to 20 parameters. Unfortunately, most detection in mass cytometry relies on lanthanide-tagged antibodies, which is ideal to detect proteins, but not other types of molecular features. To further expand the repertoire of molecular features that are detectable by mass cytometry, we developed a lanthanide-chelated, azide-containing probe that allows click-chemistry mediated labeling of target molecules. Following incorporation of the thymidine analog 5-ethynyl-2′-deoxyuridine (EdU) during DNA synthesis in S-phase of the cell cycle, we demonstrate that the probe introduced here, tagged with Terbium-159 (159Tb), reacts via copper-catalyzed azide-alkyne Huisgen cycloaddition (click-chemistry) with Edu. Thus, detection of 159Tb makes it possible to measure DNA synthesis in single cells using mass cytometry. The approach introduced here shows similar sensitivity (true positive rate) to other methods used to measure DNA synthesis in single cells by mass cytometry and is compatible with the parallel antibody-based detection of other parameters in single cells. Due to its universal nature, the use of click-chemistry in mass cytometry expands the types of molecular targets that can be monitored by mass cytometry.
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影响因子：
16.8
作者：
Bendall SC;Nolan GP;Roederer M;Chattopadhyay PK
通讯作者：
Chattopadhyay PK
DOI：
10.1126/science.1198704
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bendall SC;Simonds EF;Qiu P;Amir el-AD;Krutzik PO;Finck R;Bruggner RV;Melamed R;Trejo A;Ornatsky OI;Balderas RS;Plevritis SK;Sachs K;Pe'er D;Tanner SD;Nolan GP
通讯作者：
Nolan GP
DOI：
10.1007/978-1-4939-0944-5_7
发表时间：
2014-01-01
期刊：
EXOCYTOSIS AND ENDOCYTOSIS, 2ND EDITION
影响因子：
--
作者：
Eichelbaum, Katrin;Krijgsveld, Jeroen
通讯作者：
Krijgsveld, Jeroen
影响因子：
4.6
作者：
LONCIN, MF;DESREUX, JF;MERCINY, E
通讯作者：
MERCINY, E
影响因子：
82.9
作者：
Chattopadhyay, Pratip K.;Price, David A.;Roederer, Mario
通讯作者：
Roederer, Mario