Impact of the backbone composition on the merging quality in multi-tube cytometry experiments : flow cytometry part

Several computational approaches were developed in flow and mass cytometry to combine cell phenotypic information from different cytometric profiles. In such multi-tube cytometry experiments, investigators use antibody panels composed of cell markers specific to each panel or common to the different panels. The set of common markers is called the backbone. These cell merging can be performed at the cell or cell-population levels. However, no methodology has been proposed so far for defining the set of common markers in multi-tube cytometry experiments. Conclusion: At the cellular level, we demonstrated that the presence of lineage and differentiation/memory markers in the backbone is crucial for effective merging. Furthermore, the inclusion of functional proteins, such as cytokines, is needed to correctly predict the co-expression of other functional proteins. At the cell-population level, presence of functional protein was not mandatory to obtain effective merging. BD LSR Fortessa cytometer QC was checked before the acquisition.