Next, a flow chamber with a silanized hydrophobic coverslip was assembled and incubated with anti-digoxigenin antibodies (Roche Applied Science, 11093274910) in BRB-80 buffer for 15 minutes. The coverslip was then passivated with 1% Pluronic F-127 in BRB-80 and washed with BRB-80 buffer. Stable GMPCPP seeds were introduced into the chamber for 15-30 minutes, and unbound seeds were washed away with BRB-80 buffer. The Imaging Buffer was freshly prepared, and consisted of the BRB-80 buffer supplemented with 2 mM guanosine triphosphate (Sigma-Aldrich), 5 mg/mL bovine serum albumin, 0.08 mg/mL catalase, 0.1 mg/mL glucose oxidase, 12 mg/mL glucose, 1 mM dithiothreitol, and 0.5% β-mercaptoethanol, then centrifuged at 25,000 g at 37 °C for 15 minutes before each experiment. The polymerization mixture, containing the Imaging Buffer and 12-18 µM non-labeled tubulin, was prepared with either the tested compound in DMSO or an equivalent concentration of DMSO as a negative control. This mixture was preheated to 32 °C for 1 minute and sequentially introduced into the flow chamber.