How to make usage of the standardized EuroFlow 8-color protocols possible for instruments of different manufacturers
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
29154914
DOI
10.1016/j.jim.2017.11.007
PII: S0022-1759(17)30139-4
Knihovny.cz E-resources
- Keywords
- EuroFlow, Flow cytometry, Fluorescent spectrum, Fluorochromes, Standardization,
- MeSH
- Hematologic Neoplasms diagnosis MeSH
- Immunophenotyping instrumentation standards MeSH
- Humans MeSH
- Flow Cytometry instrumentation standards MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
A 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.
Clínica Universidad de Navarra Centro de Investigación Medica aplicada IDISNAm Pamplona Spain
Department of Immunohematology and Blood Transfusion Leiden The Netherlands
Department of Pediatric Hematology and Oncology Zabrze Medical University of Silesia Katowice Poland
Kantonsspital Aarau Aarau Switzerland
Laboratoire d'Hématologie Pôle Cellules et Tissus CHU INSERM UMR 1236 Rennes France
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