sEVs (100K pellets) derived from 1.2 × 107 oligodendrocytes were resuspended in PBS and analyzed using the NanoSight LM10 system equipped with the green laser (532 nm) and the syringe pump and NanoSight 2.3 software (Malvern) at 23°C (temperature control). The following settings were used: camera control in standard mode (camera level 16) and particle detection in standard mode (screen gain 16, detection threshold 6, and minimum expected particle size auto). Script control was used (Repeatstart, Syringeload 500, Delay 5, Syringestop, Delay 15, Capture 30, Repeat 4). Five 30-s videos were recorded, particles were tracked (batch process), and average values were formed. Particle concentration was related to the volume of culture supernatant (particles/milliliter) or the number of secreting cells (particles/cell).
Nanosight 2
Manufactured by Malvern Panalytical
Sourced in Germany, United Kingdom
The Nanosight 2.3 software is a core component of Malvern Panalytical's nanoparticle characterization technology. It provides advanced analysis capabilities for nanoparticle size, concentration, and motion. The software operates with the Nanosight instrument to enable real-time visualization and measurement of nanoparticles in liquid samples.
Automatically generated - may contain errors
7 protocols using nanosight 2
1
Characterization of Oligodendrocyte-Derived sEVs
2
Nanoparticle Analysis of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SEC-isolated EVs were analysed using the Nanosight LM10 system (camera model Hamamatsu C11440-50B/A11893-02) equipped with a 532 nm laser and a syringe pump as well as the Nanosight 2.3 software (Malvern, Herrenberg, Germany) at 23°C (temperature controlled). The following settings were used: camera control in standard mode (camera level 14), particle detection in standard mode (detection threshold 8 and minimum expected particle size auto), and script control (Repeatstart, Syringeload 500, Delay 5, Syringestop, Delay 15, Capture 30 and Repeat 4). Five videos of 30 s were recorded, particles were tracked (batch process) and average values were formed. EV samples were diluted in particle-free PBS to obtain a particle concentration of 3–10 × 108 particles/ml. Measurements were verified utilizing silica microspheres (Polysciences, Warrington, PA, USA) with a size of 100 nm as described by Gardiner [27 ].
3
Nanoparticle Characterization by NTA
4
Characterization of Magnetic Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A transmission electron microscope, JEM 1400, 120 kV (JEOL, Tokyo, Japan), was used to examine the size and morphology of the MNPs. Samples for transmission electron microscopy (TEM) were dropped onto a copper 200-mesh grid and dried. The average MNP’s size was determined by analyzing at least 50 particles using ImageJ software version 1.52 (National Institutes of Health, USA). The determination of hydrodynamic size via Nanoparticle Tracking Analysis (NTA) was carried out using a NanoSight NS500 instrument (Malvern Panalytica, Malvern, UK) equipped with an 80 mW 532 nm laser. The size distribution of the MNPs was determined using NanoSight 2.3 software (Malvern, UK). The zeta-potential of MNPs was measured via dynamic light scattering (DLS) using a ZetaSizer Nano ZS (Malvern, UK), averaging 20 runs per measurement using the Smoluchowski model. Samples for DLS were dissolved in a 10 mM KCl solution, and measurements were taken using a transparent zeta-potential cell (DTS1060C). Mössbauer spectra of 57Fe nuclei at room temperature were recorded with an MS-1104Em spectrometer (Southern Federal University, Research Institute of Physics, Rostov-on-Don, Russia) in transmission geometry with an α57Co(Rh) radiation source. Spectral analysis was performed in the Univem MS 701 program, and the relative intensities (area) of the initial spectra were determined.
5
Nanoparticle Size Analysis of Plasma Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pellets of 100,000×g resulting from 1.4 ml blood plasma were resuspended in 200 µl PBS and 1:10 dilutions were analyzed using the Nanosight LM10 system (camera model Hamamatsu C11440-50B/A11893-02) equipped with the green laser (532 nm) and the syringe pump and the Nanosight 2.3 software (Malvern, Herrenberg, Germany) at 23°C (temperature controlled). The following settings were used: camera control in standard mode (camera level 16), particle detection in standard mode (screen gain 16, detection threshold 6 and minimum expected particle size auto). Script control was used (Repeatstart, Syringeload 500, Delay 5, Syringestop, Delay 15, Capture 30 and Repeat 4). Five 30 s videos were recorded, particles were tracked (batch process) and average values were formed. Particle measurements were verified utilizing silica microspheres (Polysciences, Warrington, PA, USA) with a size of 100 and 300 nm as described in Gardiner et al. (36 ).
6
Nanoparticle Size and Concentration Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Particles were analysed using the Nanosight LM10 system (Malvern Panalytical, Herrenberg, Germany) equipped with a 532 nm laser and a syringe pump. Data were analysed using the Nanosight 2.3 software (Malvern Panalytical, Herrenberg, Germany). Particle movement was recorded in five videos, 30 s long, at a steady temperature of 23 °C. The average size and number of particles were calculated with regard to donor cell number.
7
Characterization of Magnetic Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A transmission
electronic microscope
JEM 1400, 120 kV (JEOL, Japan) was used to examine the size and morphology
of MNPs. Samples for transmission electron microscopy (TEM) were dropped
and dried onto a copper 200 mesh grid. Average MNPs’ size was
determined by analyzing at least 50 particles using ImageJ software
(National Institutes of Health, USA). Determination of hydrodynamic
size by nanoparticle tracking analysis (NTA) was carried out using
a NanoSight NS500 instrument (Malvern, UK) equipped with an 80 mW
532 nm laser. The size distribution of MNPs was determined using NanoSight
2.3 software (Malvern, UK). The zeta-potential of MNPs was measured
by dynamic light scattering (DLS) using a ZetaSizer Nano ZS (Malvern,
UK), averaging 20 runs measurement using the Smoluchowski model. Samples
for DLS were solved in 10 mM KCl solution; the measurements were taken
using a transparent zeta-potential cell (DTS1060C). Mössbauer
spectra of 57Fe nuclei at room temperature were recorded
with a MS-1104Em spectrometer (Southern Federal University, Research
Institute of Physics, Russia) in transmission geometry with a 57Co(Rh) radiation source. Spectra analysis was performed by
Univem MS program, and the relative intensities (area) of elementary
spectra were determined.
electronic microscope
JEM 1400, 120 kV (JEOL, Japan) was used to examine the size and morphology
of MNPs. Samples for transmission electron microscopy (TEM) were dropped
and dried onto a copper 200 mesh grid. Average MNPs’ size was
determined by analyzing at least 50 particles using ImageJ software
(National Institutes of Health, USA). Determination of hydrodynamic
size by nanoparticle tracking analysis (NTA) was carried out using
a NanoSight NS500 instrument (Malvern, UK) equipped with an 80 mW
532 nm laser. The size distribution of MNPs was determined using NanoSight
2.3 software (Malvern, UK). The zeta-potential of MNPs was measured
by dynamic light scattering (DLS) using a ZetaSizer Nano ZS (Malvern,
UK), averaging 20 runs measurement using the Smoluchowski model. Samples
for DLS were solved in 10 mM KCl solution; the measurements were taken
using a transparent zeta-potential cell (DTS1060C). Mössbauer
spectra of 57Fe nuclei at room temperature were recorded
with a MS-1104Em spectrometer (Southern Federal University, Research
Institute of Physics, Russia) in transmission geometry with a 57Co(Rh) radiation source. Spectra analysis was performed by
Univem MS program, and the relative intensities (area) of elementary
spectra were determined.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!