Image cytometry-based quantification protocol of human pulmonary arterial endothelial cells in lab-fabricated multichannel microfluidic devices
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.<sup>1</sup>.
Authors
- Eva Nozik‐Grayck (ORCID: https://orcid.org/0000-0002-7229-5528)
- Kurt R. Stenmark (ORCID: https://orcid.org/0000-0001-6918-6411)
- Luca Cucullo (ORCID: https://orcid.org/0000-0002-2827-7162)
- Fakhrul Ahsan (ORCID: https://orcid.org/0000-0002-1831-4708)
- Md Ibrahim (ORCID: https://orcid.org/0000-0002-2241-1500)
- Tanoy Sarkar (ORCID: https://orcid.org/0000-0001-7905-9539)
- T. D. Nguyen (ORCID: https://orcid.org/0000-0002-3862-5853)
- Marc A. Simon
- Ahmed El‐Shamy (ORCID: https://orcid.org/0000-0002-6045-5889)
- Sakib M. Moinuddin (ORCID: https://orcid.org/0000-0002-5870-7653)
- Md. Shahadat Hossain (ORCID: https://orcid.org/0000-0002-1368-7230)
- Tim Lahm
Institutions
- Pioneer (United States) (US)
- Oakland University (US)
- University of California, San Francisco (US)
- National Jewish Health (US)
- California Northstate University (US)
- VA Northern California Health Care System (US)
- University of Colorado Anschutz Medical Campus (US)
- University of Colorado Denver (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-10-22
- DOI
- https://doi.org/10.1016/j.xpro.2025.104147
- Primary Topic
- 3D Printing in Biomedical Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00