SULM – Schweizerische Union für Labormedizin | Union Suisse de Médecine de Laboratoire | Swiss Union of Laboratory Medicine

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1Institut für Labormedizin, Kantonsspital Aarau, Switzerland, 2Medizinische Immunologie, Universitätsspital Basel, Basel, Switzerland, 3Service d’hématologie, Hopitaux Universitaires de Geneve, Geneve, Switzerland, 4Universitätsklinik für Hämatologie und Hämatologisches Zentrallabor, Inselspital, Bern, Switzerland, 5Zentrum für Labormedizin, St. Gallen, 6Zentrum für Labormedizin, Luzerner Kantonsspital, Luzern, Switzerland , 7Diagnostische Hämatologie, Universitaetsspital Basel, Basel, Switzerland, 8Istituto Oncologico della Svizzera Italiana, Ospedale San Giovanni, Bellinzona, Switzerland

To generate reliable and reproducible results from flow cytometric analyses, standardization of the entire process encompassing antibody panels, instrument set-up, sample processing, data analysis, and data reporting is essential. A standardization project was proposed by the Swiss Cytometry Society (SCS) with the aim to increase comparability of immunophenotypic data among Swiss laboratories. In total, 8 centers using either BD FACSCanto II (Becton Dickinson) (n = 6) or Navios (Beckman Coulter) (n = 3) instruments participated in the project. In every center, lymphocyte subset analysis was performed on three blood samples (central send-outs) using the in house and a standardized method in parallel. For this, each of the blood samples was split into two aliquots. The first aliquot was analysed using the in house method including routine antibody panel, sample preparation, cytometer set-up, and data analysis. The second aliquot was analysed using the standardized method including uniform antibody panel (LST-12, provided by Cytognos), sample preparation, and standardized cytometer set-up using 8-peak Rainbow beads (Spherotech) according to provided standard operating protocols (SOPs) (EuroFlow). Relative numbers of Lymphocytes, B-cells, T-cells, NK-cells, CD4+ T-cells, and CD8+ T-cells were reported as % of white blood cells on provided report forms. In parallel, raw data files (FCS3/LMD) were collected for central analysis performed on “merged” samples using Infinicyt software (Cytognos). Highly comparable data were obtained at the different centers using the standardized method. Importantly, synchronized instrument set-up of BD FACSCanto II and Navios instruments enabled measurements of highly comparable data across the different platforms. Moreover, central analysis of the data allowed identification of deviations from the SOPs as well as technical issues, e.g. failure to perform correct instrument set-up. We have showed, that inter-laboratory standardization is feasible among laboratories using different instruments. However, for fully comparable inter-laboratory results, adherence to SOPs and accurate and standardized performance of sample preparation and instrument set-up without technical errors is crucial.


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