High throughput pSTAT signaling profiling by fluorescent cell barcoding and computational analysis.

ABSTRACT: Fluorescent cell barcoding (FCB) is a multiplexing technique for high-throughput flow cytometry (FCM). Although powerful in minimizing staining variability, it remains a subjective FCM technique because of inter-operator variability and differences in data analysis. FCB was implemented by combining two-dye barcoding (DyLight 350 plus Pacific Orange) with five-color surface marker antibody and intracellular staining for phosphoprotein signaling analysis. We proposed a robust method to measure intra- and inter-assay variability of FCB in T/B cells and monocytes by combining range and ratio of variability to standard statistical analyses. Data analysis was carried out by conventional and semi-automated workflows and built with R software. Results obtained from both analyses were compared to a

SUBMITTER: Tsai WL 

PROVIDER: S-EPMC6981073 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature