title: Comparable flow cytometry data can be obtained with two types of instruments, Canto II, and Navios. A GEIL study.
abstract: Flow cytometry (FC) instruments settings classically rely on local establishment of photomultipliers (PMT) voltages adapted to the measurements expected to be performed. In the era of multiparameter FC (MFC), it appears more and more desirable that comparable patterns of fluorescence are obtained in different settings. This relies on a harmonization of settings between instruments. Although this has been shown to be feasible within a given brand of flow cytometers, little information is available about broader comparisons in a given center or in a multicenter fashion. Here, we report a two-phase series of experiments first performed between a Canto II (BD Biosciences) and a Navios (Beckman Coulter) instruments in the same center. PMT values adjusted on the reference instrument (RI) Canto II were used to establish target values for PMT settings on the paired Navios practice instrument (PI). This allowed to show the good correlation of all but peaks 1 and 2 of Rainbow(®) beads between RI and PI. Using 4- or 8-color stained leukocytes, the similitude of the settings was further confirmed. A complex set of matrices was then established between five centers all equipped with both instruments. Using Bland & Altman difference comparisons for median fluorescence values, it was shown that using either Rainbow beads or CD16 stained polymorphonuclears to set-up target values on the RI CantoII, highly superimposable results could be obtained on all 9 PI. The latter were obtained using Rainbow beads or Compbeads(®) for comparisons. In summary, this two-phase study demonstrates the feasibility of different methods allowing for a robust harmonization of settings for MFC.
authors.affiliation: Hematology Laboratory, CHU St Etienne, St Priest en Jarez, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Solly
authors.forename: Lauren
authors.initials: L
authors.lastname: Rigollet
authors.forename: Lucille
authors.initials: L
authors.lastname: Baseggio
authors.forename: Julien
authors.initials: J
authors.lastname: Guy
authors.forename: Jessica
authors.initials: J
authors.lastname: Borgeot
authors.forename: Estelle
authors.initials: E
authors.lastname: Guérin
authors.forename: Agathe
authors.initials: A
authors.lastname: Debliquis
authors.forename: Bernard
authors.initials: B
authors.lastname: Drenou
authors.forename: Lydia
authors.initials: L
authors.lastname: Campos
authors.forename: Francis
authors.initials: F
authors.lastname: Lacombe
authors.forename: Marie C
authors.initials: MC
authors.lastname: Béné
mesh_terms: D002138:Calibration; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D005456:Fluorescent Dyes / Q000737:chemistry; D058851:GPI-Linked Proteins / Q000378:metabolism; D006801:Humans; D009504:Neutrophils / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism; D012015:Reference Standards; D013194:Staining and Labeling
chemical_list: C428200:FCGR3B protein, human; D005456:Fluorescent Dyes; D058851:GPI-Linked Proteins; D017452:Receptors, IgG
keywords: PMS settings; harmonization; cross-calibration; multiparameter flow cytometry