Comparison of CyTOF assays across sites: Results of a six-center pilot study.
Comparison of CyTOF assays across sites: Results of a six-center pilot study.
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DOI：
10.1016/j.jim.2017.11.008
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发表时间：
2018-03
影响因子：
2.2
通讯作者：
Maecker HT
中科院分区：
文献类型：
--
作者：
Leipold MD;Obermoser G;Fenwick C;Kleinstuber K;Rashidi N;McNevin JP;Nau AN;Wagar LE;Rozot V;Davis MM;DeRosa S;Pantaleo G;Scriba TJ;Walker BD;Olsen LR;Maecker HT
五年多来，高维质谱细胞术已被用于免疫学研究。然而，这些研究通常在一个实验室中使用一台或几台仪器进行。我们介绍了一项六中心研究的结果，该研究使用健康对照人外周血单核细胞（PBMC）和市售试剂来测试质谱流式细胞仪和操作员的研究中心内和研究中心间变异。我们使用主要研究中心生成的预染色质控品作为参考，与各研究中心染色的样本进行比较。在主要中心分析数据，包括调查两种标准化方法的效果。所有六个位点的表现相似，母细胞频率和中位信号强度（MSI）值的CV均<30%。当在每个位点使用预混合抗体混合物等分试样时，观察到背景增加，表明混合物最好新鲜制备。测试的两种标准化方法均充分用于标准化中心之间的MSI值。聚类算法揭示了轻微的差异之间的预染色和网站染色的样品，主要是由于一些抗体的背景增加。因此，我们认为多中心质谱细胞术测定是可行的。在六个国际中心中进行了一项多中心质谱细胞计数研究。预混合的抗体混合物导致少数抗体的背景更高。手动门控和聚类算法证明所有研究中心的性能充分一致。
For more than five years, high-dimensional mass cytometry has been employed to study immunology. However, these studies have typically been performed in one laboratory on one or few instruments. We present the results of a six-center study using healthy control human peripheral blood mononuclear cells (PBMCs) and commercially available reagents to test the intra-site and inter-site variation of mass cytometers and operators. We used prestained controls generated by the primary center as a reference to compare against samples stained at each individual center. Data were analyzed at the primary center, including investigating the effects of two normalization methods. All six sites performed similarly, with CVs for both Frequency of Parent and median signal intensity (MSI) values < 30%. Increased background was seen when using the premixed antibody cocktail aliquots at each site, suggesting that cocktails are best made fresh. Both normalization methods tested performed adequately for normalizing MSI values between centers. Clustering algorithms revealed slight differences between the prestained and the sites-stained samples, due mostly to the increased background of a few antibodies. Therefore, we believe that multicenter mass cytometry assays are feasible. A multicenter mass cytometry study was performed among six international centers. A premixed antibody cocktail resulted in higher background for a few antibodies. Manual gating and clustering algorithms demonstrate that all sites were adequately consistent in performance.
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影响因子：
64.5
作者：
Levine JH;Simonds EF;Bendall SC;Davis KL;Amir el-AD;Tadmor MD;Litvin O;Fienberg HG;Jager A;Zunder ER;Finck R;Gedman AL;Radtke I;Downing JR;Pe'er D;Nolan GP
通讯作者：
Nolan GP
影响因子：
4.3
作者：
Chen H;Lau MC;Wong MT;Newell EW;Poidinger M;Chen J
通讯作者：
Chen J
影响因子：
64.5
作者：
Brodin P;Jojic V;Gao T;Bhattacharya S;Angel CJ;Furman D;Shen-Orr S;Dekker CL;Swan GE;Butte AJ;Maecker HT;Davis MM
通讯作者：
Davis MM
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
影响因子：
3
作者：
Maecker, HT;Rinfret, A;Sekaly, RP
通讯作者：
Sekaly, RP