Platinum Presenter Talk at ISEV 2022 in Lyon
Dr. Clemens Helmbrecht

Principles of Colocalization-NTA

NTA is a multi-parameter technology

Nanoparticle Tracking Analysis (NTA) is well known for measuring particle size and concentration of nanoparticles suspended in water or buffer. Due to its optical design, this technology allows the user determination of zeta potential, which is the origin of ZetaView®. Furthermore, characterization of subpopulations and detection of fluorescent nanoparticles is possible with Nanoparticle Tracking Analysis. This makes NTA a multi-parameter technology.

While in classic NTA only the scatter light is analyzed, it is impossible for the user to discriminate between a bionanoparticle – the analyte – and salt precipitates from the buffer, nanobubbles or other impurities. Therefore, fluorescence detection is the basis for specific characterization of fluorescently labelled bioconjugates.

Standard (scatter) NTA Analysis
Standard (scatter) NTA Analysis

The history of the ZetaView® family

Particle Metrix’s first instrument consisted of a microscopic body where laser and camera were integrated. The electrophoretic mobility was calculated from automated tracking analysis and presented as zeta potential distribution. In 2008, the first prototype with a folded optical beam was created. This laid the foundation for future ZetaView® generations. Two years later, the first PMX-100 was launched, providing the possibility to measure size and concentration as well as zeta potential. In 2013, the PMX-110 got equipped with manual slide-in cut-off filters for fluorescence detection.
Only a few years later we had a major breakthrough with the ZetaView® TWIN. For the first time ever, Particle Metrix incorporated two collinear lasers with an automated filter slider. This groundbreaking innovation enabled the user to perform measurements in scatter and two fluorescence channels on the same sample within minutes. As a logical consequence, the ZetaView® TWIN soon had a bigger brother, the ZetaView® QUATT instrument, incorporating four wavelengths.

Colocalization-NTA with the new ZetaView® x30 series

Today, at ISEV2022 in Lyon, Particle Metrix is proud to announce the next quantum step in Nanoparticle Tracking Analysis technology:
The characterization of colocalization ratios of fluorescent particles such as vesicles, viruses or functionalized nanoparticles.

Colocalization: Illumination Volume Overlap
Illumination Volume Overlap
Colocalization: Automated Fast Switching between NTA Channels
Automated Fast Switching between NTA Channels

The quantification of colocalization ratios is ensured as two major requirements are met: First the application of new generation lasers for illumination of identical measuring volume and second ultrafast and switching time between two NTA channels, e.g. blue and red channel.

Colocalization: Double Stained HUVEC
Double Stained HUVEC
Phi6 Bacteriophage
Phi6 Bacteriophage
C-NTA data from double-stained samples
C-NTA data from double-stained samples

The evaluation of double-stained standards shows the capability of the new ZetaView® TWIN PMX-230 with colocalization feature. Bio-conjugated fluorescently labelled DNA nanoparticles and several examples of double stained extracellular vesicles and phages were analyzed with the new colocalization feature to empower the user for specific characterization of bio-nanoparticles. 

Colocalization NTA prepares the user for future applications such as phenotyping and multi-parameter characterization of biological nanoparticles.

ZetaView® x30

Next Generation
Nanoparticle Tracking Analyzer

0x consumables
5x faster switching
10x faster cleaning
12x fluorescence channels
x statistics

…and much more!

For more information on the ZetaView® or to discuss your requirements, please contact us.

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