Related Experiment Video
Updated: Aug 4, 2026
09:16
Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Published on: January 7, 2019
9.7K
High-throughput, multiplexed quantification, and sorting of single EVs at single-molecule level
Biorxiv : the Preprint Server for Biology
|November 18, 2024
Summary
We developed BDEVS, a high-throughput platform for ultra-sensitive profiling of individual extracellular vesicles (EVs) in plasma. This technology enables precise quantification and sorting of EVs, advancing cancer immunology research and diagnostics.
Area of Science:
- Biotechnology
- Nanotechnology
- Cancer Immunology
Background:
- Extracellular vesicles (EVs) are crucial in intercellular communication and disease progression.
- Current methods for EV profiling lack throughput, sensitivity, or multiplexing capabilities for complex clinical samples.
Purpose of the Study:
- To develop a novel platform for high-throughput, multiplexed, and ultra-sensitive profiling of individual EVs directly in plasma.
- To overcome limitations of existing technologies in EV analysis, enabling better understanding of tumor-immune interactions.
Main Methods:
- Developed BDEVS (Agarose Bead-based Digital Single Molecule-Single EV Sorting) platform.
- Integrated rolling circle amplification (RCA) of cleaved EV surface proteins within agarose droplets.
- Utilized flow cytometry for readout, quantification, and sorting of EVs.
Main Results:
- Achieved single molecule sensitivity and moderate multiplexing (3-plex) at high throughput (thousands of EVs/minute).
- Demonstrated a ~100x improvement in the limit of detection for EV subpopulations compared to gold standards.
- Successfully profiled and sorted EVs for immune markers (PD-L1, CD155) and a melanoma marker (TYRP-1) in plasma.
Conclusions:
- BDEVS offers unprecedented resolution for analyzing tumor-immune interactions and detecting rare EV subpopulations.
- The platform shows transformative potential for EV-based diagnostics and therapeutic monitoring in cancer immunology.
- BDEVS enables precise quantification and sorting of EVs, crucial for understanding disease heterogeneity in clinical specimens.
More Related Videos
06:12Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets
Published on: March 17, 2023
1.4K
10:16Optimization of Flow Cytometric Sorting Parameters for High-Throughput Isolation and Purification of Small Extracellular Vesicles
Published on: January 20, 2023
3.0K
Related Concept Videos
01:12Electrospray Ionization (ESI) Mass Spectrometry
Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
01:15High-Resolution Mass Spectrometry (HRMS)
The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For example, the mass of helium...
01:13Mass Analyzers: Overview
The mass analyzer is a crucial component of the mass spectrometer. In the ionization chamber, the vaporized sample is bombarded with a high-energy electron beam to generate a radical cation and further fragment into neutral molecules, radicals, and cations. A series of negatively charged accelerator plates accelerate the cations into the mass analyzer. The mass analyzer separates ions according to their mass-to-charge (m/z) ratios and then directs them to the detector. The common types of mass...
01:19Mass Analyzers: Common Types
The quadrupole mass analyzer consists of four cylindrical metal rods arranged in a diamond carrying a DC voltage and a radio-frequency AC voltage. The motion of ions through the quadrupole depends on the field strength, causing only ions of a certain m/z to resonate successfully and strike the detector at a given field strength. Though the transmission rate for these analyzers is high, the exact elemental composition of the sample is not determined because of low resolution; however, they are...
01:21Mass Spectrometry: Complex Analysis
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
01:21Tandem Mass Spectrometry
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...