The CultreCoat® Vascular Permeability Assay employs a simplified Boyden chamber design with a one micron polyethylene terephthalate (PET) membrane coated with collagen I. Ports within the migration chamber (top) allow access to the assay chamber (bottom) without dismantling the device. This design is easier to use, prevents contamination, and is adaptable for robotic high throughput systems. The assay chamber may be directly analyzed in a plate reader, eliminating transfer steps that introduce additional variability to the assay.
The confluent cell monolayer forms a barrier through cell-cell adhesion mechanisms such as junctions (adherens, tight, and gap) and desmosomes, and this barrier restricts FITC-Dextran to the inserts. Disruption of these cell-cell adhesion mechanisms creates gaps in the intercellular space that allow diffusion of FITC-Dextran into the bottom chamber, so the amount of FITC-Dextran that diffuses is related to the amount of intercellular space, which is representative of vascular permeability.