Journal: eLife
Article Title: UFMTrack, an Under-Flow Migration Tracker enabling analysis of the entire multi-step immune cell extravasation cascade across the blood-brain barrier in microfluidic devices
doi: 10.7554/eLife.91150
Figure Lengend Snippet: ( A ) Start and end time distribution of CD4 + T-cell tracks on non-stimulated or TNF- or IL-1β-stimulated primary mouse brain microvascular endothelial cells (pMBMECs). Under-flow migration tracker ( UFM Track) correctly captures increased T-cell detachment from the non-stimulated pMBMECs after 5 min when the flow is increased to physiological levels. ( B ) Quantification of CD4 + T-cell behavior in the respective categories obtained on non-stimulated and TNF- and IL-1β-stimulated pMBMECs. Error bars show the statistical error of the mean (see text for details). ( C–H ) Motility parameters of the crawling CD4 + T cells were obtained for the three endothelial stimulatory conditions. Distributions of T-cell path length ( C ), displacement ( D ), crawling speed (path/time) ( E ), migration speed (displacement/time) ( F ), variability of instantaneous T-cell crawling speed along the track (standard deviation, G ), and meandering index ( H ). ( I ) Distribution of CD4 + T-cell accelerated movement (AM) speed is a proxy metric for the T-cell adhesion to the healthy or inflamed endothelium. ( J ) Migration speed distribution of the transmigrated CD4 + T cells. Stimulation applied to the luminal side of pMBMECs affects T-cell migration at the abluminal side of pMBMECs after their transmigration. FT – T cells performed full transmigration; UT – T cells performed uncompleted transmigration. Statistical tests are performed with the Mann-Whitney U test. Statistical significance is indicated as follows: ns – Not significant (p > 0.05); * – p ≤ 0.05; ** – p ≤ 0.01; *** – p ≤ 0.001; **** – p ≤ 0.0001.
Article Snippet: 2D2 and OT-I cells were isolated respectively with magnetic CD4 + and CD8 + T-cell selection beads (EasySep, STEMCELL Technologies).
Techniques: Migration, Standard Deviation, Transmigration Assay, MANN-WHITNEY
