We used a rigorous four-color FCM assay to detect the circulating CD45dimCD34+VEGFR2+7AAD− cell subset in 10 ml of progenitor-enriched whole blood, which characteristics included (i) sampling of an important volume of blood, (ii) a pre-enrichment step, (iii) use of a viability marker (7AAD) and (iv) a multiple gating strategy (Farace et al, 2007; Taylor et al, 2009).
Supplementary Figure 7 Gate strategy used to analyze TAMs.
Fluorescence minus one (FMO) was used for gate strategy [30].
Chatham, C. H. & Badre, D. Multiple gates on working memory.
To utilize the various information carried by different gates, we investigated reconstructions using multiple gates simultaneously.
Of note, identification of tumor cells in flow cytometry normally requires the use of multiple markers and advanced gating strategies, emphasizes the strength of SOX11 as a discriminate antigen.
The use of auto debarcoding in R language demonstrates an objective, repeatable gating strategy that can be rapidly completed and which can easily handle multiple batches of experiments.
Since most background binding in MHC multimer analysis leads to signals in either one or multiple (three or more) channels (Hadrup, unpublished observation), background is reduced approximately tenfold by this gating strategy.
To identify LAIPs as specific as possible, we used the 'and' logistic gating strategy by FACSDiva software (Becton Dickinson, San Jose, CA, USA) to define final MRD population to display co-existence of multiple aberrant antigen expression if leukemic population was homogeneous.
Supplementary Figure 2 Gating strategy.
