Application Note

A new approach to nanoscale flow cytometry with theCytoFLEX nano analyzer

Nanoscale flow cytometry is a cutting-edge technique that combines the principles of flow cytometry with nanotechnology. It enables the analysis of particles at the nanoscale, providing valuable information about their size, composition, and surface properties. Nanoscale flow cytometry has numerous applications in various fields, including biology, medicine, and materials science. It allows researchers to analyze and characterize nanoparticles, extracellular vesicles (EVs), and other small particles with great precision and sensitivity. In addition to its research applications, nanoscale flow cytometry also holds promise in the development of diagnostic tools and targeted drug delivery systems. The ability to analyze nanoparticles based on their characteristics opens new possibilities for personalized medicine and nanomedicine.

While nanoscale flow cytometry is a powerful technique with numerous advantages, it also has some limitations, including:
Detection sensitivity: there are still limits to the smallest particles that can be detected reliably. Currently, flow cytometers are not designed to detect and characterize EVs smaller than 100 nm. The detection threshold for nanoparticles is influenced by factors such as background noise, autofluorescence, and the efficiency of labeling or detection probes.
Size resolution: Flow cytometry can characterize particles in the nanoscale range, but accurately resolving nanoparticles of similar sizes can be challenging. Distinguishing particles with very small size differences, such as distinguishing between 50 nm and 60 nm particles, may be difficult due to limitations in the detection and resolution capabilities of the instrument.
Sample preparation: Preparing samples for nanoparticle or EV detection can be complex. Sample preparation methods need to consider factors such as aggregation, stability, and potential alterations of particle properties during the process. Obtaining a representative and homogeneous sample can be crucial for accurate analysis.
Standardization: Nanoscale flow cytometry is a relatively new and evolving field, and the field is still working on standardizing protocols and reference materials. This can lead to variability in data collection and analysis across different laboratories, making it challenging to compare results and establish consistent methodologies.
Data analysis complexity: Flow cytometry generates complex and multidimensional datasets, which require sophisticated data analysis techniques. Analyzing and interpreting large datasets can be time consuming, requiring expertise in data processing, visualization, and statistical analysis.

 


In this application note, we will introduce the CytoFLEX nano Flow Cytometer and its workflow. This new analyzer is the first purpose-built nanoscale flow cytometer that can detect nanoparticles, such as extracellular vesicles (EVs) at least as small as 40 nm, while simultaneously performing multiparameter
fluorescent detection. Furthermore, it enables counting, characterization, and particle size definition, all within a single instrument, setting a new standard and overcoming current limitations for nanoparticle research. The CytoFLEX nano software interface, CytExpert nano, provides the sophistication to explore
the unknown at the nanoscale range while providing the ease-of-use characteristics of the CytoFLEX platform. This way, getting answers to challenging research questions becomes easier than ever for EV.
researchers.