Nanoparticle tracking analysis (NTA) was performed with the ZetaView PMX110 instrument (ParticleMetrix) and the corresponding software (ZetaView 8.04.02) was used to measure the number and the size distribution of the vesicle preparations. Samples were diluted in filtered PBS to achieve a particle concentration in between 1×107 to 1×108 particles/ml. Within this range, we confirmed the linearity of data acquisition with beads of known concentration and size (102.7±1.3 nm, Polysciences) comparable with EVs (S2 Fig ). We also determined the range of accuracy of absolute particle quantification by NTA to be in the range of 1×107 to 1×108 particles/ml, again using dilutions of calibration beads of known initial particle concentration (S2 Fig ). All samples were measured at 11 positions with three reading cycles at each position. Pre-acquisition parameters were set to a sensitivity of 75, a shutter speed of 50, a frame rate of 30 frames per second and a trace length of 15. The post-acquisition parameters were set to a minimum brightness of 20, a minimum size of 5 pixels and a maximum size of 1000 pixels.
Zetaview pmx110 instrument
Manufactured by Particle Metrix
Sourced in Germany
The ZetaView PMX110 is a particle characterization instrument that measures the size, concentration, and zeta potential of particles in a liquid suspension. It utilizes nanoparticle tracking analysis (NTA) technology to analyze the Brownian motion of particles and determine their properties.
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4 protocols using zetaview pmx110 instrument
1
Nanoparticle Tracking Analysis of Extracellular Vesicles
2
Multimodal Characterization of Nanomaterials
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Electrochemical measurements were conducted using a conventional three-electrode system in a PC-controlled Donghua DH7000 electrochemical workstation, with a glassy carbon electrode (GCE) as the working electrode, a platinum wire as the counter electrode, and an Ag/AgCl electrode (saturated KCl solution) as the reference electrode. Transmission electron microscopy (TEM), high-resolution TEM, and energy dispersive spectrometer (EDS) mapping images were obtained on a JEOL JEM-2100F operating at 200 kV. X-ray photoelectron spectroscopy (XPS) data were recorded on a Thermo Scientific ESCALAB 250Xi spectrometer with Al Kα radiation. Scanning electron microscopy (SEM) images were collected using a Hitachi S-4800 operating at 3 kV. Nanoparticle tracking analysis (NTA) measurements were performed by using a Particle Metrix ZetaView PMX110 instrument.
3
Nanoparticle Tracking Analysis of Extracellular Vesicles
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Nanoparticle tracking analysis (NTA) was performed using a ZetaView PMX110 Instrument (ParticleMetrix, Inning am Ammersee, Germany) equipped with a 405 nm laser and integrated automated fluidics. Three cycles of recordings at the ‘high’ setting (2‐s videos) were collected of each sample with camera sensitivity set at 92, shutter at 70, minimum brightness at 15, minimum size at 10 and maximum size at 5000. Temperature was set at 26°C and monitored throughout the measurements. Videos recorded for each sample were analysed with ZetaView software version 8.04.02 SP3 to determine the concentration and size of measured particles with corresponding standard error. For analysis, auto settings were used for blur, minimum track length and minimum expected particle size. The ZetaView system was calibrated with 102 nm polystyrene microbeads (ThermoFisher Scientific #3100A) prior to every analysis session. PBS (Gibco #10010‐023) was used as diluent, and to avoid contaminating particles, a fresh bottle was opened for each analysis session. At least three separate EV preparations for each condition were analysed.
4
Nanoparticle Tracking Analysis of EVs
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A ZetaView PMX110 instrument (Particle Metrix, Meerbusch, Germany) was used for nanoparticle tracking analysis (NTA) and size/concentration quantification, as previously described [17 (link)]. Briefly, 2 mL of EVs suspended in PBS with the appropriate dilution were injected into the ZetaView cell. NTA measurements were obtained at 11 positions, with two cycles at each position. During acquisition, the temperature was set to 23 °C, with a camera sensitivity of 82, 30 frames/s, and a shutter speed of 150. Commercial 100 nm polystyrene beads (Particle Metrix) were used for instrument calibration. The Zeta potential was also measured using the ZetaView PMX110 instrument.
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