English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/219674
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE
Exportar a otros formatos:

Title

How to make usage of the standardized EuroFlow 8-color protocols possible for instruments of different manufacturers

AuthorsNovákova, Michaela; Glier, Hana; Brdickova, Nadezda; Vlkova, Marcela; Santos, Ana Helena; Lima, Margarida; Roussel, Mikael; Flores-Montero, Juan; Szczepanski, Tomasz; Velden, Vincent H. J. van der; Fernandez, Paula; Mejstrikova, Ester; Burgos, Leire; Paiva, Bruno; Dongen, J. J. M. van; Orfao, Alberto ; Kalina, Tomas
KeywordsFlow cytometry
Fluorescent spectrum
Standardization
Fluorochromes
EuroFlow
Issue Date2019
PublisherElsevier
CitationJournal of Immunological 475: 112388 (2019)
AbstractA critical component of the EuroFlow standardization of leukemia/lymphoma immunophenotyping is instrument setup. Initially, the EuroFlow consortium developed a step-by-step standard operating protocol for instrument setup of ≥ 8-color flow cytometers that were available in 2006, when the EuroFlow activities started. Currently, there are 14 instruments from 9 manufacturers capable of 3-laser excitation and ≥ 8 color measurements. The specific adaptations required in the instrument set-up to enable them to acquire the standardized 8-color EuroFlow protocols are described here. Overall, all 14 instruments can be fitted with similar violet, blue and red lasers for simultaneous measurements of ≥ 8 fluorescent dyes. Since individual instruments differ both on their dynamic range (scale) and emission filters, it is not accurate to simply recalculate the target values to different scale, but adjustment of PMT voltages to a given emission filter and fluorochrome, is essential. For this purpose, EuroFlow has developed an approach using Type IIB (spectrally matching) particles to set-up standardized and fully comparable fluorescence measurements, in instruments from different manufacturers, as demonstrated here for the FACSCanto II, and Navios and MACSQuant flow cytometers. Data acquired after such adjustment on any of the tested cytometry platforms could be fully superimposed and therefore analyzed together. The proposed approach can be used to derive target values for any combination of spectrally distinct fluorochromes and any distinct emission filter of any new flow cytometry platform, which enables the measurement of the 8-color EuroFlow panels in a standardized way, by creating superimposable datafiles.
Publisher version (URL)https://doi.org/10.1016/j.jim.2017.11.007
URIhttp://hdl.handle.net/10261/219674
DOI10.1016/j.jim.2017.11.007
ISSN0022-1759
Appears in Collections:(IBMCC) Artículos
Files in This Item:
File Description SizeFormat 
howEuroFlow8.pdf1,04 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.