Comparison of CyTOF assays across sites: Results of a six-center pilot study

Research output: Contribution to journalJournal article – Annual report year: 2018Researchpeer-review

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Comparison of CyTOF assays across sites: Results of a six-center pilot study. / Leipold, Michael D.; Obermoser, Gerlinde; Fenwick, Craig; Kleinstuber, Katja; Rashidi, Narges; McNevin, John P.; Nau, Allison N.; Wagar, Lisa E.; Rozot, Virginie; Davis, Mark M.; DeRosa, Stephen; Pantaleo, Giuseppe; Scriba, Thomas J.; Walker, Bruce D.; Olsen, Lars R.; Maecker, Holden T.

In: Journal of Immunological Methods, Vol. 453, 2018, p. 37-43.

Research output: Contribution to journalJournal article – Annual report year: 2018Researchpeer-review

Harvard

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 2018, 'Comparison of CyTOF assays across sites: Results of a six-center pilot study', Journal of Immunological Methods, vol. 453, pp. 37-43. https://doi.org/10.1016/j.jim.2017.11.008

APA

Leipold, M. D., Obermoser, G., Fenwick, C., Kleinstuber, K., Rashidi, N., McNevin, J. P., ... Maecker, H. T. (2018). Comparison of CyTOF assays across sites: Results of a six-center pilot study. Journal of Immunological Methods, 453, 37-43. https://doi.org/10.1016/j.jim.2017.11.008

CBE

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. 2018. Comparison of CyTOF assays across sites: Results of a six-center pilot study. Journal of Immunological Methods. 453:37-43. https://doi.org/10.1016/j.jim.2017.11.008

MLA

Vancouver

Leipold MD, Obermoser G, Fenwick C, Kleinstuber K, Rashidi N, McNevin JP et al. Comparison of CyTOF assays across sites: Results of a six-center pilot study. Journal of Immunological Methods. 2018;453:37-43. https://doi.org/10.1016/j.jim.2017.11.008

Author

Leipold, Michael D. ; Obermoser, Gerlinde ; Fenwick, Craig ; Kleinstuber, Katja ; Rashidi, Narges ; McNevin, John P. ; Nau, Allison N. ; Wagar, Lisa E. ; Rozot, Virginie ; Davis, Mark M. ; DeRosa, Stephen ; Pantaleo, Giuseppe ; Scriba, Thomas J. ; Walker, Bruce D. ; Olsen, Lars R. ; Maecker, Holden T. / Comparison of CyTOF assays across sites: Results of a six-center pilot study. In: Journal of Immunological Methods. 2018 ; Vol. 453. pp. 37-43.

Bibtex

@article{bc093c4477e747a3b027f19e847164a6,
title = "Comparison of CyTOF assays across sites: Results of a six-center pilot study",
abstract = "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.",
keywords = "Clustering, CyTOF, Mass cytometry, Multicenter, Normalization, Standardization",
author = "Leipold, {Michael D.} and Gerlinde Obermoser and Craig Fenwick and Katja Kleinstuber and Narges Rashidi and McNevin, {John P.} and Nau, {Allison N.} and Wagar, {Lisa E.} and Virginie Rozot and Davis, {Mark M.} and Stephen DeRosa and Giuseppe Pantaleo and Scriba, {Thomas J.} and Walker, {Bruce D.} and Olsen, {Lars R.} and Maecker, {Holden T.}",
year = "2018",
doi = "10.1016/j.jim.2017.11.008",
language = "English",
volume = "453",
pages = "37--43",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Comparison of CyTOF assays across sites: Results of a six-center pilot study

AU - Leipold, Michael D.

AU - Obermoser, Gerlinde

AU - Fenwick, Craig

AU - Kleinstuber, Katja

AU - Rashidi, Narges

AU - McNevin, John P.

AU - Nau, Allison N.

AU - Wagar, Lisa E.

AU - Rozot, Virginie

AU - Davis, Mark M.

AU - DeRosa, Stephen

AU - Pantaleo, Giuseppe

AU - Scriba, Thomas J.

AU - Walker, Bruce D.

AU - Olsen, Lars R.

AU - Maecker, Holden T.

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

KW - Clustering

KW - CyTOF

KW - Mass cytometry

KW - Multicenter

KW - Normalization

KW - Standardization

U2 - 10.1016/j.jim.2017.11.008

DO - 10.1016/j.jim.2017.11.008

M3 - Journal article

VL - 453

SP - 37

EP - 43

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -