Evaluation of a Four Wavelength NTA Instrument for biomarker detection of cell line EVs

Evaluation of a Four Wavelength NTA Instrument for biomarker detection of cell line EVs

Shown with four-fold labeled EVs: Multi-wavelength NTA reduces total measurement time, minimizes the amount of sample required, and improves experimental reproducibility.

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Technology

High Throughput ZetaView® System vs High Cost of Other Methods

In addition to using Nano Particle Tracking Analysis (NTA) to measurement the size distribution and concentration of EV samples, both Microfluidic Resistive Pulse Sensing (MRPS) and the Single Particle Interference Reflectance Image Sensor (SP-IRIS) methods have been widely used as an alternative means of characterizing EVs. In this note, we relay two specific cases for using the ZetaView® NTA system to achieve relative “high-throughput” analysis of many EV samples, along estimations of throughput for MRPS & SPIRIS methods for an equivalent number of samples; further, we establish realistic estimates for the high cost of ownership for operating MRPS & SP-IRIS systems as a result of the cost of consumables as well as the substantially greater amount of time spent to run the same number of samples.

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Example of an F-NTA measurement of a bacteriophage Phi6 preparation labelled with Sybr™Gold nucleic acid stain (blue curve) and Cellbrite® Fix 640 lipid layer stain (red curve) compared to scatter-based NTA (grey curve). Purity was calculated to be 82% for dsRNA and 85% for lipid layer containing phage particles.
Application

Rapid determination of purity, integrity and titer
of viruses by Fluorescence Nanoparticle Tracking
Analysis (F-NTA) using the bacteriophage Phi6

Determination of the titer of viruses and bacteriophages is an indispensable key technology in virological research and for diagnostic purposes. Depending on the method used, the measurements are either qualitative or quantitative, very time-consuming, and do not provide information about integrity or aggregation behaviour of the virus particles. Nanoparticle Tracking Analysis with the Particle Metrix ZetaView® instrument allows the user to perform a rapid concentration determination of virus particles. Using the bacteriophage Phi6 as an example, we show how titer, purity and integrity of phage particles can be measured quickly and reliably using the fluorescence detection capability of the Particle Metrix ZetaView® instrument.

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