Fura-2 Staining
Fura-2 Staining - 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.
08:05
Staining the Cytoplasmic Ca2+ with Fluo-4/AM in Apple Pulp
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Cited by 1 •
2021
Isolated protoplasts of apple pulp cells were loaded with a calcium fluorescent reagent to detect cytoplasmic Ca2+ concentration.
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.
11:33
Microscopy and Staining: Gram, Capsule, and Endospore Staining
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2023
Source: Rhiannon M. LeVeque1, Natalia Martin1, Andrew J. Van Alst1, and Victor J. DiRita1 1 Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States of America Bacteria are diverse microorganisms found nearly everywhere on Earth. Many properties help distinguish them from each other, including but not limited to Gram-staining type, shape and arrangement, production of capsule, and formation of spores. To observe these properties, one...
FAQs
The readout changes because calcium binding shifts Fura-2’s excitation spectrum. Fluorescence is monitored near 510 nm while the dye is excited at approximately 340 and 380 nm, so the two signals reflect different calcium-sensitive spectral states. The resulting wavelength-dependent change provides the basis for estimating intracellular calcium during an immune or infection-related response.
Fura-2 AM is the form that enables cellular loading: its membrane-permeable ester can enter the cell, then intracellular esterases release the calcium-sensitive dye. This conversion is important because the measurement depends on having the active indicator inside the cell, where it can respond to calcium changes associated with stimulation or pathogen exposure.
Using the fluorescence ratio at approximately 340 and 380 nm gives a comparative signal tied to the calcium-dependent excitation shift. Researchers can therefore estimate intracellular calcium from the relationship between the two excitation responses while monitoring emission near 510 nm. This ratiometric readout is the central measurement for comparing calcium signaling across experimental responses.
A basic workflow begins by introducing Fura-2 AM to cells, allowing intracellular esterases to release the active dye, and then measuring fluorescence. The experiment compares excitation near 340 and 380 nm while recording emission near 510 nm. Researchers then use the fluorescence ratio to estimate intracellular calcium during the condition being studied.
In immunology and infection research, the method can be applied to lymphocytes, phagocytes, and infected cells. It is especially informative when the experimental question concerns how receptor stimulation or pathogen exposure changes intracellular calcium. Measurements can connect those calcium responses across different cellular contexts rather than restricting analysis to a single immune-cell type.
Calcium measurements can be interpreted alongside functional outcomes such as immune-cell activation, secretion, and cell death. This allows a study to ask whether receptor stimulation or pathogen exposure is associated only with a calcium signal or also with a broader cellular response. The approach therefore links an intracellular second-messenger event to observable consequences relevant to host responses.