Fig. 2

Evaluating neuroimmune axes using conventional 2D cultures of brain endothelial cells. a A conventional 2D model of brain endothelial cells in monoculture grown on a transwell. b Measurement of TEER using an Endohm cup chamber apparatus to evaluate BBB integrity and disruption (Axis 1), c Measurement of Pe. A fluorescent tracer is pipetted into the luminal chamber and medium is collected from the abluminal chamber over a time interval and evaluated for fluorescence. This method can be used to evaluate BBB disruption when inert tracers are used (Axis 1), and transport of labeled molecules and/or vesicles (Axis 2 and 3). d Measurement of immune cell interactions and trafficking. Leukocytes can be labeled and added to the upper chamber for evaluation of their interactions with brain endothelial cells (Axis 4). e Polarized secretions from brain endothelial cells can be measured using standard assays like ELISAs at baseline (Axis 5) and following an immune stimulus (Axis 2). f Brain endothelial cells grown on appropriate materials for imaging can be fixed and/or stained for expression/localization of tight junction proteins or vesicular changes and imaged by standard light/fluorescence microscopy or electron microscopy (Axis 1). This figure was created with BioRender