Fura-2am Loading
Fura-2am Loading - 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.
01:19
Impact Loading
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2024
Impact loading occurs when a moving object collides with a stationary structure, such as a rod with a uniform cross-sectional area fixed at one end. Under these conditions, the rod absorbs the kinetic energy from the striking object, leading to deformation and subsequent stress development. As the rod returns to its original position and reaches maximum stress, the absorbed energy, initially manifested as kinetic energy, transforms entirely into strain energy. In cases of elastic deformation,...
01:16
Eccentric Loading
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2024
Eccentric loading is a crucial concept in the study of structural engineering and mechanics, particularly when analyzing the stability and stress distribution in columns. Unlike centric loading, where the force is applied along the centroidal axis, causing uniform compression, eccentric loading occurs when a force is applied off-center. This off-center application introduces not only direct compressive stress but also bending stress, significantly influencing the column's behavior under load.
FAQs
Once the membrane-permeable indicator has crossed the plasma membrane, intracellular esterases cleave its acetoxymethyl (AM) groups. This chemical conversion releases charged Fura-2 inside the cell, so the indicator is no longer in the same membrane-permeable form. That retention step makes intracellular calcium monitoring possible over the experiment.
Calcium binding changes Fura-2’s excitation properties, producing different fluorescence responses when the indicator is excited near 340 versus 380 nm. Comparing those signals as a ratio provides the measurement used to estimate intracellular calcium concentration. The ratiometric design therefore links calcium-dependent spectral behavior to a quantitative readout of cellular signaling.
After cleavage, charged Fura-2 remains within the cell, allowing fluorescence measurements to reflect intracellular rather than merely extracellular indicator behavior. In neurons, this retained signal can be followed while calcium changes accompany activity. That feature supports observation of dynamic cellular responses instead of limiting the experiment to whether the dye entered the sample.
The workflow begins by introducing the membrane-permeable AM form to living cells and allowing it to cross the plasma membrane. Intracellular esterases then generate the charged indicator. Fluorescence is subsequently recorded with alternating excitation near 340 and 380 nm, and the ratio of those signals is used to estimate intracellular calcium during the experiment.
Neuronal activity associated with synaptic function can produce changes in intracellular calcium that the loaded indicator reports through its excitation-dependent fluorescence. By following these signals, researchers can examine calcium dynamics in relation to synaptic signaling and broader neuronal communication. The approach therefore connects optical calcium measurements with cellular events relevant to how neurons process activity.
Changes in the fluorescence ratio provide estimates of intracellular calcium that can be related to activity-dependent responses in neurons. These measurements help characterize calcium dynamics during neuronal signaling and excitability, while also supporting studies of calcium-related cellular responses. The result is a way to examine how neuronal activity or synaptic signaling is reflected in intracellular calcium behavior.