NanoSight Pro

Extracellular Vesicles (EV)

Whilst the research into the generation and functions of extracellular vesicles is still evolving, it is important to monitor and control their isolation and purification. NanoSight NTA provides quick and easy characterisation of both the size and concentration of vesicles in aqueous buffers, giving confidence in the quality of the sample used in any downstream experiments, whilst the use of fluorescence can assist in elucidating the origin subpopulations of EVs.

Virus and Vaccines

Manufacture of vaccines requires controlled production processes to ensure that the materials can be appropriately dosed and recognized by the immune system. NTA assists in the optimization of manufacturing processes by ensuring the size distribution of the vaccine is maintained. As NTA is also able to count and size viruses, it can be used as a faster alternative to titer assays.

Drug Delivery & Gene Therapy

With drug delivery and gene therapy products, 70-150 nm is the typical size of particles required to ensure effective disease targeting in the clinical. NTA is ideally suited to study the size of these particles from early stage research through to candidate screening, formulation development and clinical batch monitoring whilst the concentration measurement allows for dose determination of the final product as well as for use in in vitro and in vivo assays.

Biotherapeutics

Temperature, pH change, agitation, shear and time all impact the stability of biotherapeutic proteins, causing denaturation and aggregation which can cause loss of efficacy and the potential for an unwanted immune response. NTA provides high resolution size distributions of sub-visible aggregates in a formulation for safety and quality assurance.

Nanomaterials/colloids/toxicology

As nanomaterials are incorporated in to everyday items, they are coming to the attention of regulatory bodies. There is a requirement to fully characterize these particles, their properties and contribution in the final product. NanoSight can provide a number-based concentration for these nanomaterials and high-resolution size distributions for those working at the nanoscale in industrial, environmental and toxicological fields.

Ultrafine bubbles

In recent years work on ultrafine bubbles has been gathering momentum for use in industrial cleaning and water treatment, agriculture and food, and medical applications amongst others. Their relatively low concentration and small size are a challenge for many traditional nanoparticle characterization techniques whereas NTA is particularly adept at the detection and analysis of these bubbles.

What is Nanoparticle Tracking Analysis (NTA)?

Nanoparticle Tracking Analysis (NTA) is a technique used to analyse the size and concentration of nanoparticles in a suspension. It works by collecting video clips of the light scatter from nanoscale particles moving under Brownian motion, then tracking each particle individually to calculate size and concentration.

What is the difference between NTA and DLS?

The NanoSight Pro uses Nanoparticle Tracking Analysis (NTA) to track individual particles on a particle-by-particle basis, providing size distribution, direct particle concentration measurements, real-time visual validation, and integrated fluorescence detection with enhanced resolution for polydisperse samples and machine learning to minimise user subjectivity. The Zetasizer uses Dynamic Light Scattering (DLS) for ensemble-average size and zeta potential measurements, with minimum sample volumes from 3 uL.

How does the NanoSight work?

The NanoSight Pro illuminates nanoparticles in suspension with a laser. A high-sensitivity sCMOS camera captures each particle’s Brownian motion in real time, and the NS Xplorer software’s FTLA algorithm analyses these movements to calculate size distribution and concentration.

What particle size range does NanoSight measure?

The NanoSight Pro measures particles from 10 nm to 1,000 nm in diameter.

Can NanoSight measure particle concentration?

Yes. The NanoSight Pro measures particle concentration in the range of 10^6 to 10^9 particles/mL using a dynamic observation volume-based concentration algorithm.

What applications is NanoSight used for?

The NanoSight Pro is used for extracellular vesicle (EV) research, viral vector and vaccine manufacturing, lipid nanoparticles (LNPs) and gene therapy, biopharmaceutical aggregation detection, engineered nanomaterials, nanotoxicology, and ultrafine bubble characterisation.

What sample preparation is required for NTA?

The NanoSight Pro requires a minimum of 250 uL when using the Low Volume Flow Cell. Samples should be aqueous suspensions within the detectable concentration range of 10^6 to 10^9 particles/mL.

Can NanoSight measure exosomes?

Yes. Extracellular vesicle (EV) research which includes exosomes is listed as a primary application of the NanoSight Pro.

What factors affect NTA measurement quality?

Key factors include sample concentration, temperature (the Pro supports 10 degrees C below ambient to 70 degrees C for stability studies), particle visibility (enhanced by fluorescence detection with up to 5 filters), and flow dynamics (continuous flow delivers more statistically robust data than manual zone-based systems).

How does the NanoSight Pro differ from traditional light scattering techniques?

Unlike traditional DLS (Dynamic Light Scattering), which provides an ensemble average, the NanoSight Pro visualises and tracks individual nanoparticles in real time. This allows for direct measurement of particle-by-particle size distribution and concentration, even in polydisperse samples.

What’s new in the NanoSight Pro compared to previous NanoSight models?

NanoSight Pro delivers improved reproducibility, faster analysis, and simplified operation with new automated hardware alignment, integrated temperature control, enhanced sCMOS camera technology, and the intuitive NS Xplorer software.

How easy is it to use the NanoSight Pro?

The system is designed for ease of use with intelligent software guidance, automated alignment, and built-in self-checks that minimise operator variability, ideal for multi-user labs.

How does NTA compare with TEM and other nanoparticle characterisation techniques?

NTA provides direct measurement of individual nanoparticles, unlike DLS which measures average size of particles in suspension, and TEM which provides high-resolution images but requires a vacuum environment and is time-consuming. NTA provides higher size resolution than DLS and allows real-time visualisation in solution.

Does NTA require calibration?

NTA does not require calibration per se. Polystyrene latex standards can be routinely run to verify correct operation and check for drift. If results are outside specification or show drift, contact ATA Scientific for support.

How long does a typical NanoSight measurement take?

A complete measurement including capture and analysis can be performed in under 5 minutes per sample, with multiple replicates automated for high-throughput workflows.

Can NanoSight Pro measure fluorescently labelled nanoparticles?

Yes. The NanoSight Pro includes fluorescence modes that enable specific detection of labelled subpopulations, ideal for tracking exosomes, virus-like particles, and labelled nanocarriers in complex biological samples.

What are the maintenance requirements for NanoSight Pro?

Routine maintenance is minimal and primarily involves cleaning sample chambers and ensuring proper flushing between runs. ATA Scientific provides local technical support and training to ensure optimal system performance.

Why should I buy a NanoSight Pro?

4 reasons to choose NanoSight Pro: (1) Fast, easy-to-use and accurate size and concentration of nanoparticles. (2) Determine heterogeneity to optimise isolation and purification methods. (3) Easily confirm batch consistency for production quality assurance. (4) Fluorescence capabilities enable detection of subpopulations such as intact vesicles, common and specific biomarkers, and cargo.

Should I use DLS or NTA to measure particles below 80 nm?

DLS can measure most materials to sizes below 1 nm, and NTA is also very capable in the sub-80 nm range. The lower limit for NTA depends on the refractive index of the material: approximately 10 nm for metals, around 30 nm for polymers, and ~40 nm for liposomes. As long as particles are visible, the analysis of that image for size determination is easy and robust.

Is dilution needed for NanoSight sample preparation?

Dilution is most likely required unless the sample is very sparse to begin with. The ideal concentration range for NTA is 10^7 to 10^9 particles/mL, typically 10 to 1000x more dilute than samples used for DLS.

Can NTA detect size changes from coating a particle with surfactant or antibody?

Yes. NTA measures hydrodynamic diameter, the size of the object moving in liquid. Adding surfactants or antibodies to the particle surface affects its motion, and the reported size will shift by an amount equal to the size of the attached molecules. Size shifts as small as 3 nm have been reliably detected in coated/uncoated experiments.

Can I have a demonstration or free trial of the NanoSight Pro?

Yes. ATA Scientific offers free demonstrations and sample testing to help you evaluate the NanoSight Pro with your specific application. Contact us for a booking or complete a request a demo form.