Fura 2-am Ester
Fura 2-am Ester - 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.
04:10
Hydrolysis of an Ester - Concepts
0 Views •
2020
Triglycerides Triglycerides are triester molecules composed of a glycerol backbone that is bound to 3 fatty acids. These form the basis of many biological lipids that are found in vegetable oils and fats. Lipids that are isolated from plant material are the basis of vegetable oils, whereas lipids extracted from animals are used as lard or general sources of fats. A fatty acid molecule has a long carbon chain — between 12 and 20 carbons — with a weak organic acid at one end. Some fatty acids...
11:04
Hydrolysis of an Ester - Student Protocol
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2020
Source: Lara Al Hariri and Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA Preparation of SoapExpand Soap is prepared using a saponification reaction, where a base catalyzes the hydrolysis of three ester groups of an oil, such as coconut oil. During saponification, hydroxide ions from the base attack the carbonyl group on the oil to form a ratio of three molecules of soap to one molecule of glycerol. The resulting soap molecule is a long carbon chain, which is...
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.
FAQs
The acetoxymethyl ester groups allow the indicator to enter cells in a membrane-permeable form. Once inside, intracellular esterases cleave those groups and produce a charged Fura-2 indicator that remains in the cytoplasm. This conversion is essential because it both enables cellular loading and supports measurement of calcium changes within the cell.
Comparing fluorescence from excitation near 340 and 380 nm provides a ratiometric measurement rather than relying on a single intensity value. The resulting comparison helps reduce effects caused by differences in dye loading and illumination. Consequently, calcium-related fluorescence changes can be interpreted with less influence from these experimental variations.
After intracellular conversion, calcium binding changes the fluorescence behavior of the trapped Fura-2 indicator in a concentration-dependent manner. Imaging therefore links measured fluorescence changes to changes in intracellular Ca2+ levels. In neuronal experiments, this relationship allows activity-associated calcium signals to be quantified rather than observed only as qualitative changes in brightness.
The esterified form is membrane permeable, which allows it to enter cells. Esterase cleavage then removes the acetoxymethyl ester groups and generates a charged indicator retained in the cytoplasm. These forms therefore serve different roles in the measurement process: one supports cellular entry, while the other provides the intracellular calcium-responsive signal.
A typical sequence begins with cellular entry of the membrane-permeable ester, followed by intracellular cleavage by esterases. The resulting cytoplasmic indicator is then monitored using fluorescence excitation near 340 and 380 nm. Comparing the two fluorescence measurements provides a ratiometric readout of calcium dynamics while reducing the influence of loading and illumination differences.
In neurons, the indicator can quantify intracellular calcium changes associated with synaptic signaling and action-potential activity. These measurements connect neuronal communication or electrical activity with calcium dynamics inside the cell. The approach can therefore reveal activity-linked responses that are not captured by examining cellular behavior without an intracellular calcium readout.
Researchers can monitor calcium responses in neurons and other excitable cells while examining receptor function or responses to experimental treatments. Changes in the fluorescence-based calcium readout provide a way to compare intracellular signaling under different experimental conditions. This makes the method useful for linking treatments or receptor activation to cellular calcium responses.
Calcium is an important signal in neuronal communication and activity, so tracking its intracellular dynamics provides a direct view of signaling-related cellular responses. Fura-2-am ester supports this analysis in neurons and other excitable cells through ratiometric fluorescence measurements, helping investigators relate calcium changes to synaptic signaling, action potentials, receptor function, or treatments.