Vesicle Labeling
Vesicle Labeling - Related Videos
02:43
Quantitative Flow Cytometry to Study Labeled Protein-Phospholipid Vesicle Interactions
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2025
In this video, we demonstrate the interaction between proteins and artificial phospholipid vesicles using quantitative flow cytometry. The binding of fluorescently-labeled proteins to fluorescently-labeled phospholipid vesicles increases the mean fluorescence intensity, and this increase is detected by a flow cytometer.
08:36
FM Dyes in Vesicle Recycling
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2023
FM dyes are a class of fluorescent molecules that has found important use in studying the vesicle recycling process. By virtue of a chemical structure, these molecules can insert themselves into the outer leaflet of phospholipid bilayer membranes. After membrane insertion, they are internalized into the cell via endocytosed vesicles, and released when these vesicles recycle back to the membrane. Since, these dyes fluoresce strongly in the hydrophobic environment within membranes and weakly in...
09:01
Uptake of Fluorescent Labeled Small Extracellular Vesicles In Vitro and in Spinal Cord
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Cited by 13 •
2021
We describe a protocol to label macrophage-derived small extracellular vesicles with PKH dyes and observe their uptake in vitro and in the spinal cord after intrathecal delivery.
07:58
Labeling of Extracellular Vesicles for Monitoring Migration and Uptake in Cartilage Explants
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Cited by 5 •
2021
Here, we present a protocol to label platelet lysate-derived extracellular vesicles to monitor their migration and uptake in cartilage explants used as a model for osteoarthritis.
12:27
Measuring Peptide Translocation into Large Unilamellar Vesicles
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Cited by 9 •
2012
This protocol details a method for the quantitative measure of peptide translocation into large unilamellar lipid vesicles. This method also provides information about the rate of membrane translocation and can be used to identify peptides that efficiently and spontaneously cross lipid bilayers.
FAQs
Fluorescence changes provide a readout of membrane behavior during synaptic activity. When labels are incorporated into synaptic vesicle membranes or associated with vesicle proteins, changes in the signal can indicate vesicle exocytosis, when contents are released, and endocytosis, when membrane is retrieved. Tracking these changes helps distinguish activity-dependent release from subsequent membrane recovery.
Membrane incorporation marks the vesicle boundary and can report changes linked directly to membrane exchange during recycling. Attaching labels to vesicle-associated proteins instead follows components associated with the vesicle. These strategies emphasize different aspects of vesicle behavior, allowing researchers to examine membrane dynamics, protein localization, or both during synaptic transmission.
Stimulation activates the synaptic processes that labeled vesicles are intended to report. The resulting signal changes can show how terminals release neurotransmitters, retrieve vesicle membrane, and replenish vesicle pools. Comparing responses under different stimulation conditions helps characterize how synaptic terminals handle activity and how efficiently their vesicle supply supports neuronal communication.
A typical workflow places a label in the synaptic vesicle membrane during neuronal activity or attaches it to a vesicle-associated protein. Researchers then visualize the labeled compartments and measure changes in their signal as neurons respond to stimulation. The resulting measurements are analyzed to assess vesicle movement, exocytosis, endocytosis, and pool replenishment.
Measurements from labeled vesicles can show how synaptic terminals store and release neurotransmitters, how vesicles respond to stimulation, and how released membrane is recovered. They can also provide evidence about replenishment of vesicle pools. Together, these outcomes help researchers characterize synaptic transmission and the cellular processes supporting neuronal communication.
In neuroscience, the method supports investigations of synaptic transmission, neuronal communication, and circuit function. It can also be applied to studies of neurodevelopment and disorders involving vesicle trafficking. By linking fluorescence measurements with vesicle release, retrieval, and replenishment, researchers can examine how changes in trafficking may affect neuronal signaling.