Automated high-throughput analysis of B cell spreading on immobilized antibodies with whole slide imaging

Abstract

Automated image processing methods enable objective, reproducible and high quality analysis of fluorescent cell images in a reasonable amount of time. Therefore, we propose the application of image processing pipelines based on established segmentation algorithms which can handle massive amounts of whole slide imaging data of multiple fluorescent labeled cells. After automated parameter adaption the segmentation pipelines provide high quality cell delineations revealing significant differences in the spreading of B cells: LPS-activated B cells spread significantly less on anti CD19 mAb than on anti BCR mAb and both processes could be inhibited by the F-actin destabilizing drug Cytochalasin D. Moreover, anti CD19 mAb induce a more symmetrical spreading than anti BCR mAb as reflected by the higher cell circularity.