Protocol for live-cell calcium imaging of human lung microvascular endothelial vessel-on-a-chip model

Calcium signaling is crucial for endothelial cell homeostasis. Alterations in intracellular calcium levels due to shear stress are linked to vascular dysfunction and diseases. Here, we present a protocol to perform live calcium imaging by using a live calcium indicator on human lung endothelial cells subjected to shear stress in a commercially available microfluidic device (Ibidi Luer VI). We also outline data analysis procedures. This protocol can test altered shear stress effects in the vasculature and identify potential drug treatments. For complete information on the generation and use of this protocol, please refer to Amoakon et al.¹.