Fura-2 Fluorescence
Fura-2 Fluorescence - Related Videos
12:15
Calcium Imaging of Cortical Neurons using Fura-2 AM
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Cited by 108 •
2009
Calcium signals play a key role in many cellular processes including gene expression, survival and differentiation. Here we demonstrate how to perform calcium imaging using Fura-2 AM. Calcium imaging is a valuable tool to study the regulation of intracellular calcium in real time and its regulation of signaling cascades.
05:58
A Novel Nicotinamide Adenine Dinucleotide Correction Method for Intracellular Ca2+ Measurement with Fura-2-Analog in Live Cells
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2019
Due to the spectral overlapping of the excitation and emission wavelengths of NADH and fura-2 analogs, the signal interference from both chemicals in live cells is unavoidable during quantitative measurement of [Ca2+]. Thus, a novel online correction method of NADH signal interference to measure [Ca2+] was developed.
08:37
Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
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Cited by 3 •
2014
PAD4 is an enzyme responsible for the conversion of peptidyl-arginine to peptidyl-citrulline. Dysregulation of PAD4 has been implicated in a number of human diseases. A facile and high-throughput compatible fluorescence based PAD4 assay is described.
08:29
Assessment of Myofilament Ca2+ Sensitivity Underlying Cardiac Excitation-contraction Coupling
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Cited by 9 •
2016
This paper describes a protocol that assesses the changes of myofilament Ca2+ sensitivity during contraction in isolated cardiac myocytes from rat heart. Together with cardiac electrophysiology, systolic/diastolic cytosol Ca2+ levels and contraction/relaxation, this measurement is imperative in underpinning the mechanisms mediating cardiac excitation-contraction coupling in healthy and diseased hearts.
13:24
Fluorescence and Bioluminescence Imaging of Subcellular Ca2+ in Aged Hippocampal Neurons
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Cited by 6 •
2015
Intracellular Ca2+ remodeling in aging may contribute to excitotoxicity and neuron damage, processes mediated by Ca2+ overload. We aimed at investigating Ca2+ remodeling in the aging brain using fluorescence and bioluminescence imaging of cytosolic and mitochondrial Ca2+ in long-term cultures of rat hippocampal neurons, a model of neuronal aging.
FAQs
The two excitation bands provide complementary signals from the same indicator. Calcium binding changes how strongly Fura-2 responds near 340 and 380 nm, so the relative intensities can be compared rather than relying on a single brightness value. That ratio helps estimate free intracellular calcium while reducing sensitivity to differences in dye loading and illumination.
Emission near 510 nm serves as the readout after each excitation condition. Recording this emission at both excitation settings provides the pair of fluorescence values needed for ratiometric comparison. Keeping the emission region consistent focuses the measurement on calcium-dependent changes in excitation behavior, supporting more comparable estimates of intracellular calcium.
An altered ratio is interpreted as a change in estimated free intracellular calcium, rather than as a simple change in overall fluorescence brightness. This distinction matters because dye loading and illumination can affect absolute intensity. The ratiometric comparison helps separate those influences from calcium-related signal changes when researchers compare cellular responses.
Measurements use excitation near 340 and 380 nm and record emission near 510 nm for each condition. Researchers then compare the two fluorescence intensities to generate the calcium-related readout. This workflow preserves the paired information required for ratiometric analysis and helps make results less dependent on how much indicator is present in the cells.
Fura-2 fluorescence can be applied across medically relevant models, including neurons, muscle cells, immune cells, and cardiac tissue. In each setting, the measurement can characterize calcium-signaling responses without restricting analysis to one cell type. This broad use supports comparisons among physiological and pathological cellular behaviors in different experimental systems.
In disease-focused studies, the method can reveal altered cellular responses associated with pathology, pharmacological treatments, or toxicity. Researchers can use those response patterns to examine how cells react to interventions and to support work on diagnosis and therapeutic development. Its value lies in connecting calcium-signaling changes with clinically relevant experimental conditions.