Figure 9.

Expected outcome: target cell or cluster exhibits higher nuclear NF-κB after stimulation, analysis determines single and cluster activation thresholds

(A-B) Single cell (A) or cluster (B) stimulation before, during, and after dispensing agonist. Target cell/cluster becomes activated, neighboring cells do not respond (or respond less strongly). Green indicates reporter GFP fluorescence, purple indicates agonist concentration.

(C) Quantified individual cell responses and agonist diffusion data are combined during data analysis to compare stimulus and response on the single cell level. Results can be visualized as a plot of activation vs. location in target area, or as activation vs. agonist concentration at each location.