Home › Cell Biology › Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Cell Biology JoVE (Open Access) Citable · DOI

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

DOI: 10.3791/58731-v
What you'll learn
  • ✓ Isolate extracellular vesicles from human whole blood rapidly
  • ✓ Perform fluorescence-based staining of EV-specific markers
  • ✓ Characterize single EVs using nanoparticle-tracking analysis
Protocol

In this protocol, we describe the complete workflow for rapid isolation of extracellular vesicles from human whole blood and characterization of specific markers by fluorescence-based nanoparticle-tracking analysis. The presented results show a high level of reproducibility and can be adjusted to cell culture supernatants.

Difficulty
intermediate
Total time
~2–3 hours per sample
Biosafety
BSL-2

Steps

1
Isolate extracellular vesicles from human whole blood

Perform rapid isolation of extracellular vesicles from human whole blood using the established protocol. The isolated EVs are prepared for downstream characterization.

▶ 00:36
2
Stain isolated samples with fluorescent markers

Apply fluorescent labeling to isolated EV samples to mark specific extracellular vesicle markers. This enables fluorescence-based detection in subsequent analysis.

▶ 01:55
3
Analyze samples by nanoparticle-tracking analysis

Perform fluorescence-based nanoparticle-tracking analysis to characterize individual extracellular vesicles and quantify marker expression. Record representative data from the analysis run.

▶ 03:16
4
Interpret and evaluate characterization results

Review representative results from exosome characterization, assess reproducibility, and evaluate the quality of marker detection across analyzed samples.

▶ 06:41
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