Lamp-egfp Vesicles
Lamp-egfp Vesicles - Related Videos
11:43
Easy Measurement of Diffusion Coefficients of EGFP-tagged Plasma Membrane Proteins Using k-Space Image Correlation Spectroscopy
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Cited by 37 •
2014
This paper provides a step by step guide to the fluctuation analysis technique k-Space Image Correlation Spectroscopy (kICS) for measuring diffusion coefficients of fluorescently labeled plasma membrane proteins in live mammalian cells.
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...
06:57
Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of Eimeria that Infect Chickens
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Cited by 10 •
2015
Diagnosis of Eimeria infection in chickens remains demanding. Parasite morphology- and host pathology-led approaches are commonly inconclusive, while molecular approaches based on PCR have proven demanding in cost and expertise. The aim of this protocol is to establish loop-mediated isothermal amplification (LAMP) as a straightforward molecular diagnostic for eimerian infection.
08:46
Studying Synaptic Vesicle Pools using Photoconversion of Styryl Dyes
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Cited by 18 •
2010
FM dyes have been of invaluable help in the understanding of synaptic dynamics. FMs are normally followed under the fluorescent microscope during different stimulation conditions. However, photoconversion of FM dyes combined with electron microscopy allows the visualization of distinct synaptic vesicle pools, among other ultrastructure components, in synaptic boutons.
10:03
Development of Leishmania Species Strains with Constitutive Expression of eGFP
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2023
Here, we describe the methodology used for generating L. panamensis and L. donovani strains expressing the gene for eGFP as a stable integrated transgene using the pLEXSY system. Transfected parasites were cloned by limiting dilution, and clones with the highest fluorescence intensity in both species were selected for further use in drug screening assays.
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The LAMP-EGFP fusion places EGFP fluorescence on vesicle membranes, so the fluorescent signal marks the boundary and movement of individual endolysosomal compartments in living cells. Because the label remains associated with the membrane, microscopy can follow vesicle trajectories rather than only measuring overall lysosomal abundance. This makes changes in transport and compartment behavior visible over time.
Microtubules provide the routes along which LAMP-EGFP vesicles move through axons and dendrites. Tracking fluorescence along these neuronal processes can show whether compartments are transported through extended cellular regions and where their movement changes. This spatial information is important because lysosomal trafficking must connect distant neuronal compartments with sites requiring cargo degradation or maintenance.
Delivery, docking, and fusion represent successive stages in how endolysosomal compartments reach and interact with neuronal destinations. Observing these events helps distinguish a transport problem from a defect in the later contact or fusion step. Such distinctions can clarify how lysosomal function supports cargo degradation and synaptic maintenance, rather than treating all vesicle accumulation as the same cellular defect.
A live-cell workflow uses neurons expressing the LAMP-EGFP fusion protein and fluorescence microscopy to observe labeled vesicles over time. Imaging can follow their movement through axons and dendrites, then examine delivery, docking, and fusion within neuronal compartments. The resulting recordings provide dynamic observations of trafficking that fixed observations cannot capture as directly.
Imaging can provide evidence about vesicle movement, distribution through axons and dendrites, delivery to neuronal compartments, and interactions involving docking or fusion. These observations help characterize lysosomal trafficking and reveal whether endolysosomal behavior is coordinated with neuronal maintenance. Comparing these dynamic outcomes across conditions can expose altered compartment handling without relying only on endpoint fluorescence.
They are useful when researchers need to examine lysosomal trafficking in neurons, including processes related to cargo degradation, synaptic maintenance, or responses to injury. Their dynamics can also expose cellular defects associated with neurodegenerative disease and impaired endolysosomal function. In these settings, fluorescence-based tracking links visible transport behavior with broader changes in neuronal compartmental health.