Breaking the Boundaries of Extracellular Vesicle Detection

Beckman Coulter CytoFLEX nano 

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Get ready to break the boundaries of EVs detection! Go Deeper !

Extracellular vesicles are key players in intercellular communication, playing a significant role in the development of immune responses, tissue regeneration, tumor growth, and more. However, their analysis is complicated by the limitations of traditional cytometry in detecting EVs smaller than 100 nm .

With the CytoFLEX nano, lower the limits of detection, so you can achieve more.

A new approach to nanoscale flow cytometry with the  BECKMAN COULTER CytoFLEX nano   

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, 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. 
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Beckman Coulter CytoFELX nano flowcytometer EVs detection

The CytoFLEX nano 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. 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.

What Makes CytoFLEX nano Unique?

Built on the same principles as the CytoFLEX platform, the CytoFLEX nano combines the capabilities of various methods into a single platform, ensuring the detection of nanoparticles as small as 40 nm, analysis of individual EV markers even with low expression (<10 antigen copies), and the ability to provide standardized, fast, and accurate analysis without using multiple systems.


Source Beckman Coulter - Figure 1. The size of different cells and extracellular vesicles. The CytoFLEX Nano Flow Cytometer can detect, characterize, and determine the size of extracellular vesicles as small as 40 nm

Key Advantages and Features

    Sensitivity and Multi-Channel Detection
    6 fluorescent channels and 5 side scatter channels allow detailed analysis of even the smallest particles.
    Increased laser power and specially designed optical excitation paths collect more light, enhancing the efficiency of the analysis.
    Data Purity and Contamination Minimization
    Automated cleaning tools quickly detect and eliminate contamination, minimizing the risk of sample loss to <1%.
    The daily quality control process detects noise from nano- and micro-bubbles and triggers automatic purging and cleaning.
    Optimized Optical and Fluidic Systems
    Reduced sample and sheath fluid flow rates enable working with micro-samples, ensuring high accuracy of results.
    Sample collection with a small-volume piston pump ensures stability and accuracy of counting (>90%).
    Specialized sheath fluid, combined with a variable 5-nm filter, minimizes background noise.
    Ease of Use
    The intuitive CytExpert software allows easy system setup and adaptation to the needs of users at any level.
    Process automation reduces time and simplifies sample preparation. 


The CytoFLEX nano combines everything in only one instrument, enabling count, characterization, and size determination 

TOTAAL TESTBEHEER

Particle size determination by flow cytometry requires calibration using an appropriately characterized reference particle paired with a calibration method. Latex polystyrene particles are the tool of choice for this task due to their NIST-traceable nature and reliably consistent manufacturing. Through the calibration method taught to each CytoFLEX nano flow cytometer customer, data will be reliably reportable in absolute units for method comparison and correlation.


COUNT 

The most robust counting methods rely on highly controlled volumetric delivery of samples rather than hand loading samples with the implied variability. The CytoFLEX nano flow cytometer allows users to customize their sample handling so that the right amount of sample is used for the results needed.


 CHARACTERIZE

The ability to describe nanoscale particles requires best-in-class sensitivity. To make this possible, the CytoFLEX nano flow cytometer has completely redesigned the optics and fluidics system to maximize the detection of even the smallest amounts of fluorescence across 6 detection channels and excited by 4 collinear lasers.  

Explore the full picture of your EV experiment with the CytoFLEX nano Flow Cytometer. 

This purpose-built flow cytometer provides greater sensitivity, consistent instrument performance, and flexibility to study your sample.

Sensitivity

With an unparalleled size sensitivity from 1µm to 40nm* and 10nm resolution,** the CytoFLEX nano flow cytometer enables the detection of smaller EV populations and their low-abundance cargo. 


Flexibility

Open up your research with 6 fluorescent channels and 5 side scatter channels. Gather all the necessary data with fewer constraints.

 

Maximum Consistency

Maintaining contamination-free, robust instrument performance is critical when working with small size particles. Obtain reproducible results you can trust with >90% volumetric counting accuracy and <1% carryover between samples thanks to automated QC controls. 

Familiarity

Built on the same principles as the CytoFLEX platform, the CytoFLEX nano flow cytometer features a similar footprint and user-friendly software interface.  




Interested in learning how to advance your Extracellular Vesicle research?


Discover how the BECKMAN COULTER CytoFLEX nano Flow Cytometer shatters the previous limits of detection capabilities, arming researchers with data they previously would have never been able to access.


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When choosing a flow cytometry instrument, there are three key factors to consider:

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Counting, determining the size of, and characterizing extracellular vesicles (EVs) gives researchers deeper insights into cell and organ health and disease profiles. However, EVs can be challenging to analyze, owing to their small size and heterogeneity.
The need for standardization
Until now, scientists have had to use multiple techniques to accurately count, characterize and determine the size of EVs, to get the insights they need. As a result, workflows that are time-consuming and have poor repeatability are often required. 

extracellular Vesicle secreting cell

 

Webinar: First Experiences with the CytoFLEX nano

This webinar features Alfonso Blanco, Director of Flow at University College Dublin, who will present preliminary results that researchers who study extracellular vesicles (EVs) have obtained using the CytoFLEX nano Flow Cytometer.  
This webinar explores the limits of flow cytometry technology and outline how the CytoFLEX nano flow cytometer is designed to help address these limitations in a robust and consistent system able to standardize results between labs. Our expert speaker, Alfonso Blanco, discuss the advantages of using fluorescence combined with multiple scatters for further characterization of nanoparticles and extracellular vesicles. View the replay   

Press Review: What They Say About Recent Advances and Insights in Extracellular Vesicle Research

In the article, "Developments in Extracellular Vesicle (EV) Research Present a Promising Future", published in Technology Network - July 2024, Matthew Goff, Beckman Coulter Product Manager, talks about the following topics:

- Traditional challenges in EV research
- Emergence of new technologies
- The importance of workflow automation
- How nanoscale technology is accelerating EV research 

"....With more clarity in results, EVs offer advanced diagnostic, prognostic, theranostic and therapeutic procedures, while simultaneously reducing the invasive nature of sample collection. Monitoring disease progression or creating custom therapeutic models is incredibly powerful but it requires specificity in analytical methods. This is finally available for a variety of researchers. 
Updating this workflow is really an exciting moment in research....”

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Beckman Coulter Life Sciences Revolutionizes Nanoparticle Analysis With Launch of CytoFLEX Nano Flow Cytometer.

Article  published in Technology Network : March 28, 2024

“...The CytoFLEX nano Flow Cytometer shatters the previous limits of detection capabilities, arming researchers with data they previously would have never been able to access.” This can open new doors for researchers and enable breakthrough discoveries in our battle against a myriad of diseases. Extracellular vesicles can be found everywhere, and now researchers finally have a tool to see what has always been right in front of them. We listened to the needs of our customers to create an analytical crossroads by increasing sensitivity through enhanced fluidics, optics, and electronics......”

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Extracellular vesicles (EVs) are small membrane-bound particles released by cells into the extracellular environment. They play important roles in intercellular communication and are involved in various physiological and pathological processes. EVs are classified into different subtypes based on their biogenesis and size, including exosomes, microvesicles, and apoptotic bodies. Exosomes or small EVs typically range in size from 30 to 150 nm. They are formed through the inward budding of multivesicular bodies (MVBs) within the cell, which then fuse with the plasma membrane, releasing the exosomes into the extracellular space. Exosomes contain various bioactive molecules, such as proteins, lipids, nucleic acids (DNA, RNA), and microRNAs, which can be transferred to target cells, influencing their function and behavior. Microvesicles, also known as large EVs or microparticles or ectosomes, are larger than exosomes, ranging from 100 to 1000 nm in size. Unlike exosomes, microvesicles are formed by the outward budding and shedding of the plasma membrane directly. They also carry a diverse cargo of proteins, lipids, and nucleic acids, and can transfer these molecules to recipient cells.
Apoptotic bodies are the largest EVs, typically ranging from 1 to 5 µm. They are released from dying cells during the process of programmed cell death (apoptosis). Apoptotic bodies contain cellular fragments, organelles, and nuclear material, and are recognized and engulfed by phagocytic cells to facilitate their clearance.

EVs have gained significant attention in recent years due to their potential as biomarkers for disease diagnosis and prognosis, as well as their roles in cell-to-cell communication and their therapeutic applications. Researchers are studying EVs in various biological fluids, including blood, urine, and cerebrospinal fluid, to gain insights into their cargo and functions.
Technologies such as nano flow cytometry, electron microscopy, and molecular profiling techniques like RNA sequencing and proteomics are used to study and characterize EVs. Understanding EV biology, cargo, and functions holds great promise for advancing our knowledge of cellular communication and their potential applications in diagnostics, therapeutics, and regenerative medicine. Currently, EV analysis is critical and challenging. Isolation and purification methods can suffer from low yield, contamination from other particles, and difficulties in standardization. Heterogeneity of EV populations in terms of size, cargo, and biogenesis complicates their study. EVs can exhibit diverse biological activities and functions depending on their cellular origin and cargo. However, deciphering the specific functions and mechanisms of action of EVs in different contexts is still a challenge. The functional heterogeneity of EVs requires more comprehensive characterization and standardized functional assays. Researchers in the field of EVs are actively working to address these limitations by developing improved isolation techniques, standardizing protocols, and advancing our understanding of EV biology. As the field progresses, overcoming these challenges will help unlock the full potential of EV research and its applications in various biomedical areas. One of the biggest limitations is the need for multiple techniques to accurately count, characterize and determine the size of EVs, resulting in time-consuming and laborious workflows with poor repeatability.