ArticleAnalytical chemistry2023
Efficient Labeling of Vesicles with Lipophilic Fluorescent Dyes via the Salt-Change Method.
Article in Analytical chemistry, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Cardiac immunity and heart repair: Mechanism, challenge, and future direction.Chinese medical journal · 2026Review
- Integrated multi-omics profiling of gut bacterial extracellular vesicles links cargo composition to host transcriptional responses.bioRxiv : the preprint server for biology · 2026Article
- Endogenous Labelling of Extracellular Vesicles and Image Capture of Their Interactions With Acceptor Cells.Chembiochem : a European journal of chemical biology · 2026Article
- Macromolecule-Loaded Hybrid Extracellular Vesicles via Ionic Lipid-Based Post-Loading for Intracellular Delivery: Functional Evaluation for Neurodegenerative Therapy.Journal of extracellular biology · 2026Article
- Three-Dimensional Mapping of Facial Motor Neurons with Tissue Clearing for Facial Nerve Nucleus of Guinea Pig via Long Distance Retrograde Tracing.Acta histochemica et cytochemica · 2026Article
- Adipose Tissue-Derived Extracellular Vesicle MicroRNAs: Diagnostic Biomarkers for the Pathophysiology Associated with Obesity.Precision chemistry · 2025Review
- Human umbilical cord mesenchymal stem cells-derived extracellular vesicles as a therapeutic approach to ameliorate bladder injury in animal models of radiation cystitis.Stem cell research & therapy · 2025Article
- Affinity-Based Isolation and One-Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Inhalable Mucociliary-On-Chip System Revealing Pulmonary Clearance Dynamics in Nanodrug Delivery.ACS nano · 2025Article
- Incorporation of recombinant proteins into extracellular vesicles by Lactococcus cremoris.Scientific reports · 2025Article
- Impact of Culture Duration on the Properties and Functionality of Yeast-Derived Extracellular Vesicles.Biomaterials research · 2025Article
- Advancing membrane biology: single-molecule approaches meet model membrane systems.BMB reports · 2025Review
- Bacterial Extracellular Vesicles (BEVs) Derived FromInternational journal of biomaterials · 2025Article
- Extracellular Vesicles: Advanced Tools for Disease Diagnosis, Monitoring, and Therapies.International journal of molecular sciences · 2024Review
- Target specification and therapeutic potential of extracellular vesicles for regulating corneal angiogenesis, lymphangiogenesis, and nerve repair.The ocular surface · 2024Review
- Nanoparticle Tracking Analysis: An Effective Tool to Characterize Extracellular Vesicles.Molecules (Basel, Switzerland) · 2024Review
- Emerging Designs and Applications for Biomembrane Biosensors.Annual review of analytical chemistry (Palo Alto, Calif.) · 2024Review
- Fluorescent Chiral Quantum Dots to Unveil Origin-Dependent Exosome Uptake and Cargo Release.ACS applied bio materials · 2024Article
- Factors to consider before choosing EV labeling method for fluorescence-based techniques.Frontiers in bioengineering and biotechnology · 2024Review
- Assessing In Vivo Bacterial Extracellular Vesicle (BEV) Biodistribution Using Fluorescent Lipophilic Membrane Stains.Methods in molecular biology (Clifton, N.J.) · 2024Article
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Authors and funding
12 authors.
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Abstract
Fluorescent labeling allows for imaging and tracking of vesicles down to single-particle level. Among several options to introduce fluorescence, staining of lipid membranes with lipophilic dyes provides a straightforward approach without interfering with vesicle content. However, incorporating lipophilic molecules into vesicle membranes in an aqueous solution is generally not efficient because of their low water solubility. Here, we describe a simple, fast (<30 min), and highly effective procedure for fluorescent labeling of vesicles including natural extracellular vesicles. By adjusting the ionic strength of the staining buffer with NaCl, the aggregation status of DiI, a representative lipophilic tracer, can be controlled reversibly. Using cell-derived vesicles as a model system, we show that dispersion of DiI under low-salt condition improved its incorporation into vesicles by a factor of 290. In addition, increasing NaCl concentration after labeling induced free dye molecules to form aggregates, which can be filtered and thus effectively removed without ultracentrifugation. We consistently observed 6- to 85-fold increases in the labeled vesicle count across different types of dyes and vesicles. The method is expected to reduce the concern about off-target labeling resulting from the use of high concentrations of dyes.
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