Fura-2 Calcium Imaging
Fura-2 Calcium Imaging - 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.
08:43
Calcium Imaging in Neurons
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2023
Calcium ions play an integral role in neuron function: They act as intracellular signals that can elicit responses such as altered gene expression and neurotransmitter release from synaptic vesicles. Within the cell, calcium concentration is highly dynamic due to the presence of pumps that selectively transport these ions in response to a variety of signals. Calcium imaging takes advantage of intracellular calcium flux to directly visualize calcium signaling in living neurons.This video begins...
08:10
In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice
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Cited by 28 •
2008
To understand network dynamics of microcircuits in the neocortex, it is essential to simultaneously record the activity of a large number of neurons . In-vivo two-photon calcium imaging is the only method that allows one to record the activity of a dense neuronal population with single-cell resolution .
10:30
Prolonged Incubation of Acute Neuronal Tissue for Electrophysiology and Calcium-imaging
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Cited by 17 •
2017
Once removed from the body, neuronal tissue is greatly affected by environmental conditions, leading to eventual degradation of the tissue after 6 - 8 h. Using a unique incubation method, which closely monitors and regulates the extracellular environment of the tissue, tissue viability can be significantly extended for >24 h.
03:41
In Vitro Calcium Imaging in Short-Term and Long-Term Rat Hippocampal Cultures
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2025
Source: Calvo-Rodríguez, et al. Fluorescence and Bioluminescence Imaging of Subcellular Ca2+ in Aged Hippocampal Neurons. J. Vis. Exp. (2015). This video demonstrates the use of fluorescence imaging to measure cytosolic calcium levels in short-term and long-term rat hippocampal cultures. By comparing young and aged neurons, it highlights how increased L-type voltage-gated calcium channel density and prolonged activation in aged neurons result in calcium overloading, a hallmark of neuronal aging.
FAQs
The two-wavelength ratio makes the readout responsive to calcium-dependent spectral changes rather than relying on a single fluorescence intensity. Calcium binding alters Fura-2’s excitation spectrum, so comparing fluorescence excited near 340 nm with that excited near 380 nm provides a ratiometric indication of cytosolic Ca2+. This is especially useful for following dynamic signaling responses.
Fura-2 AM serves as the form that permits dye loading into cells. Once inside, intracellular esterases cleave the AM group, converting the loaded compound into the intracellular indicator used for calcium measurements. This conversion links the loading step to later calcium-sensitive fluorescence, allowing the assay to monitor responses in immune or infected cells.
The time-dependent ratio reveals how cytosolic Ca2+ changes after a receptor is stimulated or a pathogen is encountered. In immunology and infection studies, the resulting trace can be related to immune-cell activation, host responses, or cellular dysfunction. Examining rapid fluxes helps connect an external trigger with intracellular signaling behavior.
A basic workflow begins by loading cells with membrane-permeable Fura-2 AM, allowing intracellular esterases to cleave the dye, and then measuring fluorescence with excitation near 340 and 380 nm. Researchers compare the two excitation signals as cells receive receptor stimulation or pathogen exposure. The resulting ratio tracks the associated cytosolic calcium response.
The approach can be applied to lymphocytes, phagocytes, and infected cells, making it relevant across distinct stages of host defense and infection biology. Measurements after receptor stimulation or pathogen exposure can show whether these cells generate calcium fluxes and can help compare activation-associated signaling with responses linked to cellular dysfunction.
These measurements can help clarify how receptor signals and pathogen exposure are connected to intracellular calcium behavior. By tracking rapid fluxes in relevant cell types, researchers can investigate signaling pathways, characterize cellular dysfunction, and identify potential targets for immunological research. The method therefore links fluorescence measurements with mechanisms underlying host responses and infection-associated changes.