EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally compa...

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Main Authors:	Kalina, T, Flores-Montero, J, van der Velden, V H J, Martin-Ayuso, M, Böttcher, S, Ritgen, M, Almeida, J, Lhermitte, L, Asnafi, V, Mendonça, A, de Tute, R, Cullen, M, Sedek, L, Vidriales, M B, Pérez, J J, te Marvelde, J G, Mejstrikova, E, Hrusak, O, Szczepański, T, van Dongen, J J M, Orfao, A
Format:	Online
Language:	English
Published:	Nature Publishing Group 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437409/

id	pubmed-3437409
recordtype	oai_dc
spelling	pubmed-34374092012-09-10 EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols Kalina, T Flores-Montero, J van der Velden, V H J Martin-Ayuso, M Böttcher, S Ritgen, M Almeida, J Lhermitte, L Asnafi, V Mendonça, A de Tute, R Cullen, M Sedek, L Vidriales, M B Pérez, J J te Marvelde, J G Mejstrikova, E Hrusak, O Szczepański, T van Dongen, J J M Orfao, A Special Report The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally comparable results among different laboratories. This required the selection of optimal combinations of compatible fluorochromes and the design and evaluation of adequate standard operating procedures (SOPs) for instrument setup, fluorescence compensation and sample preparation. Additionally, we developed software tools for the evaluation of individual antibody reagents and antibody panels. Each section describes what has been evaluated experimentally versus adopted based on existing data and experience. Multicentric evaluation demonstrated high levels of reproducibility based on strict implementation of the EuroFlow SOPs and antibody panels. Overall, the 6 years of extensive collaborative experiments and the analysis of hundreds of cell samples of patients and healthy controls in the EuroFlow centers have provided for the first time laboratory protocols and software tools for fully standardized 8-color flow cytometric immunophenotyping of normal and malignant leukocytes in bone marrow and blood; this has yielded highly comparable data sets, which can be integrated in a single database. Nature Publishing Group 2012-09 2012-09-05 /pmc/articles/PMC3437409/ /pubmed/22948490 http://dx.doi.org/10.1038/leu.2012.122 Text en Copyright © 2012 Macmillan Publishers Limited http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Kalina, T Flores-Montero, J van der Velden, V H J Martin-Ayuso, M Böttcher, S Ritgen, M Almeida, J Lhermitte, L Asnafi, V Mendonça, A de Tute, R Cullen, M Sedek, L Vidriales, M B Pérez, J J te Marvelde, J G Mejstrikova, E Hrusak, O Szczepański, T van Dongen, J J M Orfao, A
spellingShingle	Kalina, T Flores-Montero, J van der Velden, V H J Martin-Ayuso, M Böttcher, S Ritgen, M Almeida, J Lhermitte, L Asnafi, V Mendonça, A de Tute, R Cullen, M Sedek, L Vidriales, M B Pérez, J J te Marvelde, J G Mejstrikova, E Hrusak, O Szczepański, T van Dongen, J J M Orfao, A EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
author_facet	Kalina, T Flores-Montero, J van der Velden, V H J Martin-Ayuso, M Böttcher, S Ritgen, M Almeida, J Lhermitte, L Asnafi, V Mendonça, A de Tute, R Cullen, M Sedek, L Vidriales, M B Pérez, J J te Marvelde, J G Mejstrikova, E Hrusak, O Szczepański, T van Dongen, J J M Orfao, A
author_sort	Kalina, T
title	EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
title_short	EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
title_full	EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
title_fullStr	EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
title_full_unstemmed	EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols
title_sort	euroflow standardization of flow cytometer instrument settings and immunophenotyping protocols
description	The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally comparable results among different laboratories. This required the selection of optimal combinations of compatible fluorochromes and the design and evaluation of adequate standard operating procedures (SOPs) for instrument setup, fluorescence compensation and sample preparation. Additionally, we developed software tools for the evaluation of individual antibody reagents and antibody panels. Each section describes what has been evaluated experimentally versus adopted based on existing data and experience. Multicentric evaluation demonstrated high levels of reproducibility based on strict implementation of the EuroFlow SOPs and antibody panels. Overall, the 6 years of extensive collaborative experiments and the analysis of hundreds of cell samples of patients and healthy controls in the EuroFlow centers have provided for the first time laboratory protocols and software tools for fully standardized 8-color flow cytometric immunophenotyping of normal and malignant leukocytes in bone marrow and blood; this has yielded highly comparable data sets, which can be integrated in a single database.
publisher	Nature Publishing Group
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437409/
_version_	1611555292909142016

EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

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