Calcium Green-5n Fluorescence
Calcium Green-5n Fluorescence - Related Videos
03:13
Mitochondrial Calcium Uptake Assay: A Plate Reader-Based Method for Measuring Calcium Uptake in Isolated Mitochondria
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2025
This video describes a plate reader-based assay to measure mitochondrial calcium uptake in isolated mitochondria. The assay analyzes the fluorescence signal of a calcium-sensitive fluorescent dye to study the kinetics of mitochondrial calcium uptake and calcium overload.
02:48
Fluorescence Imaging of Calcium Dynamics at Neuromuscular Junctions in the Diaphragm of a Transgenic Mouse
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2025
Source: Heredia, D. J., et. al. Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm. J. Vis. Exp. (2018) This video demonstrates fluorescence imaging of calcium dynamics at neuromuscular junctions (NMJs) in a transgenic mouse expressing genetically encoded calcium indicators. A NJM is identified by fluorescently tagged acetylcholine receptors in the diaphragm with the phrenic nerve. An image splitter is used to visualize multiple signals...
05:59
Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos
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Cited by 24 •
2015
A method for fluorescent staining of fixed biological material with the specific DNA label methyl green is described. Methyl green is used in a diluted aqueous solution and is very resistant to photobleaching. Its far-red emission allows for deep specimen imaging, making it particularly adequate for whole embryos.
11:49
Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay
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Cited by 26 •
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The here described fluorescence-based calcium mobilization assay is a medium-throughput reverse pharmacology screening system for the identification of functionally activating ligand(s) of orphan G protein-coupled receptors (GPCRs).
08:21
Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor
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Cited by 9 •
2017
This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution.
FAQs
The dye’s chelating site binds free calcium, and that binding produces an increase in fluorescence. Researchers measure the resulting intensity change and relate it to calcium availability through calibration. Because the signal depends on measurement conditions, fluorescence should be interpreted as a calibrated indicator of free calcium rather than as an unqualified concentration measurement.
Low affinity allows the indicator to remain informative in calcium-rich environments where higher-affinity dyes may not resolve additional changes accurately. This property extends the measurable range toward elevated calcium levels, making the technique particularly useful for examining calcium storage and release in systems that maintain substantial calcium availability.
Calibration conditions are central to interpreting the signal because fluorescence intensity reflects calcium availability only under an appropriate measurement relationship. The relevant calcium range also matters: the indicator is especially suited to environments with high calcium levels. Matching the dye’s response characteristics to the system helps researchers distinguish meaningful calcium changes from an uninterpreted intensity variation.
The principal distinction is the calcium range each indicator can resolve effectively. Calcium Green-5N’s lower affinity makes it better suited to elevated calcium environments, whereas higher-affinity indicators may be less suitable when calcium levels are high. Choosing between them therefore depends on the expected calcium conditions and whether the study focuses on storage, release, or signaling changes.
Researchers select a calcium-containing biological system, monitor fluorescence as calcium availability changes, and interpret intensity changes using suitable calibration conditions. The measurement can then be organized around storage, release, or rapid signaling events. This workflow connects the optical signal to calcium behavior while keeping the system’s calcium range central to experimental interpretation.
The method can be applied to studies of calcium storage and release in organelles, cells, and membrane systems. Its low-affinity response is especially relevant when these compartments or systems contain high calcium levels. Researchers can use the resulting fluorescence changes to examine how calcium availability varies across structures or during intracellular events.
In biological techniques, Calcium Green-5N fluorescence supports investigation of calcium homeostasis, transport, and physiology. Measurements can reveal changes associated with calcium storage or release and can follow rapid intracellular signaling events. These applications help connect calcium availability with the behavior of organelles, cells, and membrane systems under study.