Nanoparticle Tracking Analysis (NTA) is widely used for measuring nanoparticle size and concentration, but common challenges like photobleaching and low signal-to-noise ratios can compromise data quality, especially in fluorescence mode. In a recent study, we evaluated the performance of the Vesiculabs CD63 GFP vesicle standard, focusing on fluorophore stability utilizing low laser power.

How the Envision Solves NTA Fluorescence Issues

• Lower Laser Power: With a highly sensitive imaging system and precise laser power adjustments, the Envision uses significantly less power during fluorescence measurements—drastically reducing photobleaching.
• Enhanced Light Sheet Alignment: By reshaping the laser beam to match the exact focal volume of the observation optics, we can ensure accurate particle tracking and minimize background noise.
• Improved Fluorophore Stability: Thanks to these innovations, fluorescent particles retain their stability longer during measurements, allowing for significantly higher recovery in fluorescence vs scatter mode—far exceeding the 10% that’s typical in other NTA systems.

Vesiculab’s CD63-GFP Vesicle Standard

The Envision was able to run the CD63-GFP EV sample using a laser power of only 4 mW, significantly lower than the 50 mW typically used in other systems. This adjustment allowed the GFP-labeled vesicles to remain stable for a full 60 seconds, ensuring continuous particle tracking without the rapid loss of fluorescence.

EV CD63-GFP Under Scatter	EV CD63-GFP Under Fluorescence

During the test, the vesicles appeared bright and stable against a dark background, producing clear, high-quality data throughout the measurement. This is crucial for applications like exosome and virus detection, where even minor fluctuations in data quality can impact research outcomes. The prolonged fluorescence stability during measurement enabled precise data collection and minimized photobleaching, which is a persistent issue in legacy NTA systems.

To see more data on this sample, visit Products 2 – Hyperion Analytical

Conclusion

The data collected from the Vesiculab’s CD63 GFP vesicle standard underlines the Envision’s ability to maintain fluorophore stability with low laser power, offering more reliable and precise measurements. These improvements make the Envision a powerful tool for fluorescence-based NTA, especially in sensitive applications such as extracellular vesicle research.

For more details on the CD63 GFP sample, you can visit Vesi-Ref CD63-GFP – Vesiculab