Nanosight lm10 nanosizer

Particle size distributions were evaluated for the ID93–liposome mixture as well as each component alone (at the same concentration) using a NanoSight LM10 nanosizer with a 40 mW 410 nm laser (Malvern Instruments) and a Hamamatsu Orca Flash 2.8 CMOS camera (Hamamatsu Photonics KK, Hamamatsu, Japan). Before and after each measurement the sample cell was rinsed with three cell volumes of Millipore Milli-Q (Billerica, MA, USA) ultrapure water. Samples were diluted 1:10⁶ or 1:10⁵ to obtain a final particle concentration of approximately 5 × 10⁸ particles/mL. Appropriate dilution factors were determined empirically. Samples were diluted in ultrapure Milli-Q water in two or three steps. Each formulation was diluted four times, independently, to account for dilution error. One milliliter of diluted formulation was loaded into a 1-mL syringe and infused into the sample chamber at 15 μL/minute. Four consecutive 60-second videos were recorded for each dilution. Shutter and gain settings were optimized for each sample. The camera histogram gating was adjusted to maximize sensitivity. Data analysis was performed in NanoSight NTA 2.3 (Malvern Instruments) in batch mode.