Work

Image cytometry-based quantification protocol of human pulmonary arterial endothelial cells in lab-fabricated multichannel microfluidic devices

Year: 2025

Type: article

Abstract: Here, we present a protocol for automated quantification and viability analysis of pulmonary arterial cells (PACs) using a cellular image cytometer. We describe steps for obtaining PACs from the Pulmonary Hypertension Breakthrough Initiative, culturing and seeding them on microfluidic chips, and integrating chips into 6-well plates. Cells are stained with Hoechst 33342 and propidium iodide for live/dead analysis. Automated imaging and software-assisted quantification standardize workflows and broaden applications in pulmonary vascular research. For complete details on the use and execution of this protocol, please refer to Al Hilal et al. 1 • Steps for automated imaging and counting of cells on microfluidic chips • Instructions for staining cells with Hoechst and propidium iodide dyes • Procedures for positioning microfluidic devices in standard 6-well plate formats • Guidance for setting up imaging protocols and software-assisted quantification Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for automated quantification and viability analysis of pulmonary arterial cells (PACs) using a cellular image cytometer. We describe steps for obtaining PACs from the Pulmonary Hypertension Breakthrough Initiative, culturing and seeding them on microfluidic chips, and integrating chips into 6-well plates. Cells are stained with Hoechst 33342 and propidium iodide for live/dead analysis. Automated imaging and software-assisted quantification standardize workflows and broaden applications in pulmonary vascular research.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104147