Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment

Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner

Flow cytometric analysis was completed with a BD Accuri flow cytometer as follows: Sample volume was set to 100 µL and flow rate was set to 'fast'. Unstained and single-stained controls were used to set gates. Forward scatter (pulse area) vs. side scatter (pulse area) plots were used to gate out non-cellular debris. Forward scatter (pulse area) vs. forward scatter (pulse height) plots were used to gate out doublets. The appropriate fluorescent channels were used to determine stained vs. unstained cells. The gates were placed using unstained control samples. Single-stain controls were tested and showed there was no overlap/bleed-through between the fluorophores. Final analysis indicated the quantity of leukocytes (CD45-positive cells) and neutrophils (Ly6G-positive cells) in BAL samples.