“…Labelled pseudovirions previously incubated with 100 µM CJF-III-288 (38), and 50 µg/ml of each 17b and A32 monoclonal antibodies, or with the same concentrations of CJF-III-288, 17b, A32, plus mAb 246D, for 1 hour at room temperature, were immobilized on streptavidin-coated quartz slides and imaged on a custom-built wide-field prism-based TIRF microscope (61, 64). Imaging was performed in phosphate-buffered saline (PBS) pH ∼7.4, containing 1 mM trolox (Sigma-Aldrich, St. Louis, MO, USA), 1 mM cyclooctatetraene (COT; Sigma-Aldrich, St. Louis, MO, USA), 1 mM 4-nitrobenzyl alcohol (NBA; Sigma-Aldrich, St. Louis, MO, USA), 2 mM protocatechuic acid (PCA; Sigma-Aldrich, St. Louis, MO, USA), and 8 nM protocatechuate 3,4-deoxygenase (PCD; Sigma-Aldrich, St. Louis, MO, USA) to stabilize fluorescence and remove molecular oxygen.…”