Fura-2 Am
Fura-2 Am - 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.
08:29
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.
01:13
meta-Directing Deactivators: –NO2, –CN, –CHO, –CO2R, –COR, –CO2H
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2023
All meta-directing substituents are deactivating groups. These substituents withdraw electrons from the aromatic ring, making the ring less reactive toward electrophilic substitution. For example, the nitration of nitrobenzene is 100,000 times slower than that of benzene because of the deactivating effect of the nitro group. The first step in an electrophilic aromatic substitution is the addition of an electrophile to form a resonance-stabilized carbocation. The energy diagrams for the...
01:20
2° Amines to N-Nitrosamines: Reaction with NaNO2
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2023
Secondary amines react with nitrous acid to form N-nitrosamines, as depicted in Figure 1. Nitrous acid, a weak and unstable acid, is formed in situ from an aqueous solution of sodium nitrite and strong acids, such as hydrochloric acid or sulfuric acid, in cold conditions. In the presence of an acid, the nitrous acid gets protonated. The subsequent loss of water results in the formation of the electrophile known as nitrosonium ion. Figure 1. The nitration reaction of secondary amines The...
FAQs
Calcium binding changes Fura-2 fluorescence differently when the dye is excited at approximately 340 versus 380 nm. Measuring both signals and using their ratio allows calcium-related changes to be distinguished from some variation in dye loading or illumination. This paired-excitation principle helps track intracellular calcium changes more reliably than a single fluorescence measurement.
Intracellular esterases remove the acetoxymethyl ester groups after entry, converting the loaded compound into charged Fura-2 that remains inside the cell. This intracellular retention is important because the calcium-sensitive form must be present within the neuron to report changes associated with stimulation or signaling. The processing step therefore links membrane entry to usable intracellular measurements.
Ratiometric readouts help separate calcium-dependent fluorescence changes from experimental variation caused by unequal dye loading or changes in illumination. Rather than relying on one absolute intensity, the experiment compares signals obtained under the two excitation conditions. In neuronal imaging, this makes intracellular calcium changes easier to interpret when cells do not contain identical amounts of indicator.
First, the membrane-permeant form is introduced to living cells, including neurons. Intracellular esterases then release the charged indicator, after which fluorescence is measured with excitation near 340 and 380 nm. Calcium-dependent differences between those signals are compared as a ratio, producing a readout of intracellular calcium changes during the selected cellular condition or stimulation.
It is useful when the research question concerns calcium changes during neuronal stimulation, synaptic activity, or intracellular signaling. These experiments can visualize activity-associated calcium influx or mobilization in living neurons. The resulting fluorescence measurements connect cellular activity with calcium dynamics, helping investigators examine processes that influence neuronal excitability and intracellular signaling.
Changes detected with this indicator can be interpreted in relation to calcium influx, calcium mobilization, cellular excitability, and calcium homeostasis. A change in the measured fluorescence ratio may indicate altered calcium handling during activity or signaling. The method therefore provides functional calcium information that helps characterize how neurons respond to defined cellular conditions.