NTA measurements were performed with a NanoSight LM20 (NanoSight, Amesbury, United Kingdom), equipped with a sample chamber with a 640-nm laser and a Viton fluoroelastomer O-ring. The samples were injected in the sample chamber with sterile syringes (BD Discardit II, New Jersey, USA) until the liquid reached the tip of the nozzle. All measurements were performed at room temperature except the live monitoring protein heat stress measurements (see section below).
The software used for capturing and analyzing the data was the NTA 2.0 Build 127. The samples were measured for 40 s with manual shutter and gain adjustments. The “single shutter and gain mode” was used to capture the monodisperse polystyrene beads, the 60/100 nm beads mixture, the liposomes, the TMC particles and the protein aggregates. The “extended dynamic range mode,” which splits the capture video into two videos with independent shutter and gain settings, was used for all the other mixtures of monodisperse polystyrene beads, the PLGA particles and the insulin aggregates. Three measurements of the same sample were performed for all the polystyrene beads and six measurements for the polymer nanoparticles and protein aggregates. The error bars displayed on the NTA graphs were obtained by the standard deviation of the different measurements of each sample. The mean size and SD values obtained by the NTA software correspond to the arithmetic values calculated with the sizes of all the particles analyzed by the software.
The software used for capturing and analyzing the data was the NTA 2.0 Build 127. The samples were measured for 40 s with manual shutter and gain adjustments. The “single shutter and gain mode” was used to capture the monodisperse polystyrene beads, the 60/100 nm beads mixture, the liposomes, the TMC particles and the protein aggregates. The “extended dynamic range mode,” which splits the capture video into two videos with independent shutter and gain settings, was used for all the other mixtures of monodisperse polystyrene beads, the PLGA particles and the insulin aggregates. Three measurements of the same sample were performed for all the polystyrene beads and six measurements for the polymer nanoparticles and protein aggregates. The error bars displayed on the NTA graphs were obtained by the standard deviation of the different measurements of each sample. The mean size and SD values obtained by the NTA software correspond to the arithmetic values calculated with the sizes of all the particles analyzed by the software.
