Fm Dye Labeling
Fm Dye Labeling - Related Videos
06:48
Antibody Labeling with Fluorescent Dyes Using Magnetic Protein A and Protein G Beads
0 Views •
Cited by 3 •
2016
The on-bead method for labeling antibodies with small molecules enables labeling of a small amount of antibodies directly from cell media. This method is compatible with amine and thiol chemistry, and can handle multiple samples in parallel, manually or using automated platforms.
04:47
Retrograde Labeling of Drosophila Motor Neurons Using Fluorescent Lipophilic Dyes
0 Views •
2025
Source: Inal, M. A. et. al., Retrograde Tracing of Drosophila Embryonic Motor Neurons Using Lipophilic Fluorescent Dyes. J. Vis. Exp. (2020)This video demonstrates the method of retrograde labeling of Drosophila motor neurons using lipophilic dyes. The dye enters the axon, travels retrogradely, and labels the neurons green, allowing detailed visualization of their morphology and projections under a confocal microscope.
07:42
Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging
0 Views •
Cited by 33 •
2008
This video shows techniques for labeling of human embryonic stem cells and mesenchymal stem cells with fluorescent dyes. This technique can be used for an in vivo tracking of transplanted stem cells with optical imaging and for histopathological correlations with fluorescence microscopy.
10:55
Retrograde Labeling of Retinal Ganglion Cells in Adult Zebrafish with Fluorescent Dyes
0 Views •
Cited by 3 •
2014
We introduce an efficient method to retrograde label retinal ganglion cells (RGCs) in adult zebrafish.
14:43
Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes
0 Views •
Cited by 14 •
2008
A simple and specific method was demonstrated for fluorescent labeling and enhanced detection of cell surface proteins without a fractionation step. Differential abundance in cell surface proteins was analyzed using two-dimensional (2-D) electrophoresis and Ettan™ DIGE technology.
FAQs
Amphipathic styryl dyes partition into lipid environments, so their fluorescence increases when they associate with membranes. During endocytosis, labeled membrane is retrieved into vesicles, retaining dye within the vesicular compartment. This creates a fluorescence signal that reflects membrane uptake rather than merely the presence of dye in the surrounding solution, allowing recycling events to be followed in living cells.
When labeled vesicles fuse with the plasma membrane, their dye is exposed to the external environment and released from the recycling compartment. The associated fluorescence therefore declines after exocytosis. Monitoring this decrease provides an indirect measure of vesicle release and helps connect stimulus-driven membrane fusion with the timing and extent of synaptic or secretory activity.
An increase in fluorescence is associated with dye uptake as membrane is internalized during endocytosis, whereas a decrease indicates dye loss during exocytosis. Comparing these changes helps resolve successive stages of membrane recycling rather than treating fluorescence as a single static measurement. The pattern can therefore indicate whether a cellular response is linked to uptake, release, or both.
FM dye labeling reports membrane movement through changes caused by endocytic trapping and subsequent exocytic release. A fluorescence signal is therefore interpreted in relation to vesicle uptake and discharge, not just membrane location. This distinction makes the technique useful for studying dynamic trafficking and stimulus-dependent secretion in living cells, where membrane recycling changes over time.
The dye is allowed to associate with the plasma membrane, after which endocytosis traps it in vesicles. Researchers then monitor fluorescence during cellular stimulation and subsequent membrane trafficking. Uptake is inferred from dye accumulation, while release is assessed from fluorescence loss. This sequence links observed signal changes to the corresponding phases of vesicle recycling.
Fluorescence measurements can provide evidence for vesicle uptake, release, and recycling during neuronal activity. Because the signal changes as labeled membrane enters and exits vesicles, researchers can compare trafficking under different stimulation conditions. The resulting observations help relate cellular activity to synaptic membrane dynamics and can reveal altered patterns associated with impaired synaptic function.
The method is particularly useful when investigators need to monitor membrane recycling in living cells without directly disrupting the trafficking process. In neuronal studies, it supports analysis of neurotransmission and stimulus-dependent secretion. It can also help investigate defects in synaptic function by showing whether activity is associated with altered vesicle uptake, release, or recycling.