Journal: bioRxiv
Article Title: Neutrophils exhibit distinct migration phenotypes that are modulated by transendothelial migration
doi: 10.1101/2024.10.17.618860
Figure Lengend Snippet: (A) Schematic of pocket device for in vitro 3D directed neutrophil migration assays. (B) Sample image of cells migrating in a pocket device with Trackmate cell tracking results overlayed. Inset shows the time sequence for a chemotaxing cell, with tracked trajectory overlayed in green. Scale bar, 1000 µm. (C) Probability density maps of aggregated trajectory endpoints are shown for dHL-60 cells in each of the four conditions with representative sample tracks overlayed. Sample track color saturation increases with time. The dashed red reference circle superimposed on each inset figure represents unbiased random motion. fMLP concentration increases linearly in the positive y direction. Black arrows show the large angle turns. Scale bar, 20 µm. (D) Population average chemotactic index (CI). (E) Population average mean cell speeds in four treatment conditions in the presence of fMLP gradient. Error bars represent 95% confidence interval. All pairwise conditions showed statistically significant differences besides those labeled n.s. (not significant) in (D-E). **** P < 0.0001 according to the one-way ANOVA Tukey’s test. (F) Mean squared displacements (MSDs) versus distance separation along cell trajectories, r 2 (Δ s ). (G) Tangent vector autocorrelations versus distance traveled, ( Ctt ((Δ s )), for f MLP condition, of four treatment conditions. The inset represents the enlarged portion of the initial decay. (H) Migratory persistence length, L P . Values reflect the characteristic length of the slow decaying component of the double exponential fit, with error bars calculated via bootstrapping (see Methods). **** P < 0.0001 according to the Brown-Forsythe ANOVA test.
Article Snippet: Based on the presence of two distinct decay regimes evident in semi-log plots of C tt , the migratory persistence length for each condition was estimated by fitting C tt to a double exponential decay model, expressed as for each treatment condition (lsqcurvefit, MATLAB).
Techniques: In Vitro, Migration, Cell Tracking Assay, Sequencing, Concentration Assay, Labeling, Plasmid Preparation
