High-Throughput Mass Cytometry Staining for Immunophenotyping Clinical Samples

被引：15

作者：

Thrash, Emily M. ^{[1
]}

Kleinsteuber, Katja ^{[1
]}

Hathaway, Emma S. ^{[1
]}

Nazzaro, Matthew ^{[1
]}

Haas, Eric ^{[2
]}

Hodi, F. Stephen ^{[1
]}

Severgnini, Mariano ^{[1
]}

机构：

[1] Dana Farber Canc Inst, Ctr Immuno Oncol, Boston, MA 02215 USA

[2] Dana Farber Canc Inst, Longwood Med Area CyTOF Core, Boston, MA 02215 USA

来源：

STAR PROTOCOLS | 2020年 / 1卷 / 02期

关键词：

D O I：

10.1016/j.xpro.2020.100055

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

As mass cytometry (MC) is implemented in clinical settings, the need for robust, validated protocols that reduce batch effects between samples becomes increasingly important. Here, we present a streamlined MC workflow for high-throughput staining that generates reproducible data for up to 80 samples in a single experiment by combining reference sample spike-in and palladium-based mass-tag cell barcoding. Although labor intensive, this workflow decreases experimental variables and thus reduces technical error and mitigates batch effects.

引用

页数：28

共 7 条

[1] Automated Data Cleanup for Mass Cytometry [J].

Bagwell, Charles Bruce ;

Inokuma, Margaret ;

Hunsberger, Benjamin ;

Herbert, Donald ;

Bray, Christopher ;

Hill, Beth ;

Stelzer, Gregory ;

Li, Stephen ;

Kollipara, Avinash ;

Ornatsky, Olga ;

Baranov, Vladimir .

CYTOMETRY PART A, 2020, 97 (02) :184-198

[2] Separation, banking, and quality control of peripheral blood mononuclear cells from whole blood of melanoma patients [J].

Holland, Martha ;

Cunningham, Rachel ;

Seymour, Lake ;

Kleinsteuber, Katja ;

Cunningham, Amy ;

Patel, Tara ;

Manos, Michael ;

Brennick, Ryan ;

Zhou, Jun ;

Hodi, F. Stephen ;

Severgnini, Mariano .

CELL AND TISSUE BANKING, 2018, 19 (04) :783-790

[3] A Beginner's Guide to Analyzing and Visualizing Mass Cytometry Data [J].

Kimball, Abigail K. ;

Oko, Lauren M. ;

Bullock, Bonnie L. ;

Nemenoff, Raphael A. ;

van Dyk, Linda F. ;

Clambey, Eric T. .

JOURNAL OF IMMUNOLOGY, 2018, 200 (01) :3-22

[4] Standardization and quality control for high-dimensional mass cytometry studies of human samples [J].

Kleinsteuber, Katja ;

Corleis, Bjoern ;

Rashidi, Narges ;

Nchinda, Nzuekoh ;

Lisanti, Antonella ;

Cho, Josalyn L. ;

Medoff, Benjamin D. ;

Kwon, Douglas ;

Walker, Bruce D. .

CYTOMETRY PART A, 2016, 89A (10) :903-913

[5] A Modified Injector and Sample Acquisition Protocol Can Improve Data Quality and Reduce Inter-Instrument Variability of the Helios Mass Cytometer [J].

Lee, Brian H. ;

Kelly, Geoffrey ;

Bradford, Shermineh ;

Davila, Melanie ;

Guo, Xinzheng V. ;

Amir, El-Ad David ;

Thrash, Emily M. ;

Solga, Michael D. ;

Lannigan, Joanne ;

Sellers, Brian ;

Candia, Julian ;

Tsang, John ;

Montgomery, Ruth R. ;

Tamaki, Stanley J. ;

Sigdel, Tara K. ;

Sarwal, Minnie M. ;

Lanier, Lewis L. ;

Tian, Yuan ;

Kim, Cheryl ;

Hinz, Denise ;

Peters, Bjoern ;

Sette, Alessandro ;

Rahman, Adeeb H. .

CYTOMETRY PART A, 2019, 95 (09) :1019-1030

[6] Development of an 8-color antibody panel for functional phenotyping of human CD8+cytotoxic T cells from peripheral blood mononuclear cells [J].

Patel, Tara ;

Cunningham, Amy ;

Holland, Martha ;

Daley, John ;

Lazo, Suzan ;

Hodi, F. Stephen ;

Severgnini, Mariano .

CYTOTECHNOLOGY, 2018, 70 (01) :1-11

[7] Stabilizing Antibody Cocktails for Mass Cytometry [J].

Schulz, Axel R. ;

Baumgart, Sabine ;

Schulze, Julia ;

Urbicht, Marie ;

Gruetzkau, Andreas ;

Mei, Henrik E. .

CYTOMETRY PART A, 2019, 95A (08) :910-916

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