Particle Metrix, developers of versatile particle characterization solutions for the life sciences, report on the work of the Burger Laboratory at Ottawa Hospital Research Institute. The Lab is studying the role of microparticles in the pathogenesis of diabetic vascular and kidney disease.
Dr Dylan Burger leads a research group at Ottawa Hospital’s Research Institute. Their research focuses on understanding the biological role of membrane vesicles (specifically exosomes and microparticles/microvesicles) in vascular and renal disease. In particular, they are interested in the impact of disease on the rate of formation and molecular makeup of these vesicles.
They chose the ZetaView system from Particle Metrix for microparticle characterization because it allows them to obtain both quantitative information on the number of particles in a particular preparation as well as basic physical information (i.e. size and zeta potential). This has utility for in assessment of vesicles as biomarkers but also in quality control in vesicle preparations. When coupled with fluorescence detection, it also allows for immunophenotyping of vesicles to determine cell origin or vesicle content.
Talking about his experiences in particle characterization prior to the use of Zetaview, Dr Burger said “We have previously used electron microscopy, dynamic light scattering, flow cytometry, tunable resistive pulse sensing along with other nanoparticle tracking analysis systems. Now, with ZetaView, we have a system which delivers high detection sensitivity over a range of samples. These may be detected at low concentration levels and we have the ability to visualize fluorescent particles as well as to measure zeta potential in real time. The automation of the system (auto-alignment and autofocusing) expedites analysis and also allows for better across-lab standardization. Low maintenance needs allow us to use this system on a daily basis. Last but not least, we like that the system has a very small footprint so can be placed anywhere in a standard biomedical laboratory.”
After successfully showing C-NTA of double-stained MSC-EVs in our previous study (https://bit.ly/43GG0Yk) and confirming
the results with image flow cytometry, this note describes C-NTA of multi-stained MSC-EVs using a PMX-430 QUATT NTA
system with four lasers.
In this new application note we describe a quick determination of the physical titer of coronaviruses by fluorescence-based Nanoparticle Tracking Analysis and specific spike antibody recognition.
In this report, we describe the quantification of colocalization ratios on double-stained MSC-derived EVs using fluorochrome conjugated antibodies against the
cell surface antigens CD9 and CD41, which also serve as EV marker proteins.
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.
Along with generally accepted methods are some “tricks of the trade” such as the addition of Tween® or BSA; however, some of those additions are problematic.
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.
Headquarter
Support
