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Published on: April 25, 2013
Image cytometry-based quantification protocol of human pulmonary arterial endothelial cells in lab-fabricated
Md Ibrahim1, Sakib M Moinuddin2, Md Shahadat Hossain2
1Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, California Northstate University, Elk Grove, CA 95757, USA; Masters of Pharmaceutical Sciences Program, College of Graduate Studies, California Northstate University, Elk Grove, CA 95757, USA; Oncovask Therapeutics LLC, Elk Grove, CA 95758, USA.
None:
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.
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