Multi-site reproducibility of a human immunophenotyping assay in whole blood and peripheral blood mononuclear cells preparations using CyTOF technology coupled with Maxpar Pathsetter, an automated data analysis system
| Autor: | Shahab Asgharzadeh, Justin Lyberger, Yujiao Zhao, Greg Stelzer, Benjamin C. Hunsberger, Christopher M. Bray, Stephen K.H. Li, Azucena Gómez-Cabrero, Michael A. Strausbauch, Donald J. Herbert, Gregory K. Behbehani, Jose C. Villasboas, Ofir Goldberger, Gregory Kelly, Adeeb Rahman, Kevin D. Pavelko, Charles Bruce Bagwell, Thirumahal Selvanantham, Margaret Inokuma, Beth L. Hill, Ruth R. Montgomery, Hsiaochi Chang |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Data Analysis Canada Laboratory Proficiency Testing Histology Population Biology Peripheral blood mononuclear cell Article Pathology and Forensic Medicine Immunophenotyping Pattern Recognition Automated Automated data 03 medical and health sciences 0302 clinical medicine Immune system Humans Mass cytometry education Whole blood Automation Laboratory education.field_of_study Reproducibility Blood Cells Reproducibility of Results Cell Biology Reference Standards Flow Cytometry United States 030104 developmental biology 030220 oncology & carcinogenesis Leukocytes Mononuclear Biomedical engineering |
| Zdroj: | Cytometry B Clin Cytom |
| ISSN: | 1552-4957 |
| Popis: | High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping. |
| Databáze: | OpenAIRE |
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