Fig. 1
From: Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function
Mini-XPerT–simultaneous measurements of barrier function and cellular contractile forces in a high-throughput format. A Silicone-based elastic substrates (Young’s Modulus = 0.3 kPa or 3 kPa) were prepared in a 96-well plate format. Embedded in the substrate are fluorescent beads (diameter = ~400 nm; shown in red) whose displacement enables the computation of monolayer contractile forces (shown as white arrows). Ligated to the substrate is biotinylated collagen I (shown as black triangles) whose binding with FITC-avidin (shown in green) enables the identification of paracellular gaps. B Representative gap images for every experimental condition, C corresponding traction force maps, and D-E averaged values. While gap formation and contraction were increased with thrombin (1 U/ml, 30 min) it was significantly reduced by pre-treatment for 20 min with the ROCK inhibitor, Y-27632 (5 µM). In D, plotted is the gap area normalized to the vehicle group. In E, on a well-by-well basis, we quantified the ratio of the average monolayer contraction after vs. before stimulation. We further normalized this ratio to the vehicle group. Each treatment group comprises of measurements performed over n = 7–16 individual wells of a 96-well plate. Plotted is the mean and standard error. *indicates p < 0.05 while ns indicates p > 0.05
