Single-vesicle Tracking
Single-vesicle Tracking - Related Videos
02:35
Single-Cell Culture of Bacteria Within Giant Vesicles
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2025
Source: Morita, M., et al. Bacterial Cell Culture at the Single-cell Level Inside Giant Vesicles. J. Vis. Exp. (2019).This video demonstrates a method for culturing and observing individual bacterial cells encapsulated inside giant vesicles anchored to a supported lipid bilayer. The system enables stable, long-term imaging of single-cell growth dynamics in a confined, physiologically relevant microenvironment.
08:39
Tracking Single Proteins in Lipid Bilayers Using Fluorescence Microscopy
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2025
This article provides a detailed description of how to create samples for single-protein tracking in solid-supported lipid bilayers. It also explains a straightforward fluorescence microscope with single-molecule sensitivity and a fast frame rate. Finally, we outline the procedure for extracting single-protein trajectories.
09:16
Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
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Cited by 17 •
2019
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.
11:28
3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
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Cited by 10 •
2014
In this video protocol we track - at high speed and in three dimensions - fluorescently labeled lysosomes within living cells, using the orbital tracking method in a modified two-photon microscope.
07:29
Tracking miRNA Release into Extracellular Vesicles using Flow Cytometry
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2023
Here, we describe a straightforward protocol that enables in vitro assessment of the abundance of fluorescently labeled microRNAs to study the dynamics of microRNA packaging and export into extracellular vesicles (EVs).
FAQs
Trajectory analysis follows changes in an individual vesicle’s position over time and relates those movements to defined stages of its cycle. Researchers can therefore quantify movement through the cell, arrival at a release site, fusion-associated release, and subsequent recycling rather than treating vesicles as one averaged population. This separation helps connect dynamic behavior with specific cellular processes.
These components provide distinct regulatory points for vesicle behavior. Molecular motors and cytoskeletal tracks are associated with cargo movement, whereas presynaptic machinery regulates events at the release site, including secretion. By resolving individual trajectories and release events, the method helps researchers examine how these systems coordinate neuronal cargo transport with neurotransmitter release.
Defined cellular conditions make vesicle behavior easier to compare across experiments or neuronal states. Because the method quantifies transport, docking, fusion, and recycling at the level of individual particles, researchers can determine whether a condition changes a particular stage rather than only producing a general change in fluorescence or cell-wide secretion. This supports more specific mechanistic interpretation.
The approach resolves release events from individual vesicles, allowing vesicle behavior to be examined in relation to neurotransmitter secretion. This provides a direct cellular link between presynaptic vesicle dynamics and synaptic transmission. In neuroscience, that connection can clarify how release machinery operates and how altered vesicle behavior may contribute to differences in neuronal communication.
Researchers first label vesicles or their cargo with fluorescent probes in living cells. They then acquire time-lapse microscopy images under defined cellular conditions, reconstruct the trajectories of individual particles, and quantify behaviors such as transport, docking, fusion, and recycling. The resulting measurements describe vesicle dynamics rather than relying only on population-level fluorescence signals.
Fluorescent probes make selected vesicles or their cargo visible during time-lapse imaging. This labeling enables researchers to follow individual particles and reconstruct their paths through living cells. Because the signal identifies the cargo being observed, the resulting trajectories can be analyzed for transport and release-related behaviors under the cellular conditions established for the experiment.
Single-vesicle Tracking is useful when researchers need to compare vesicle behavior between healthy and disease-related neuronal conditions. Measurements of transport, docking, fusion, recycling, and individual release events can indicate which aspects of vesicle handling differ between states. These comparisons help relate altered presynaptic dynamics to changes in neuronal cargo movement and neurotransmitter secretion.