Calcium Am
Calcium Am - Related Videos
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...
08:14
Assessing T-cell Receptor-Induced Calcium Influx Using a Calcium Indicator Dye
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2025
This video analyzes T-cell receptor-induced calcium influx in T-cells. Anti-CD3 antibodies activate the T-cell signaling pathways, leading to an increase in intracellular calcium ions that bind with Indo-1 dye. In flow cytometry, a spectrum shift from the calcium-free state to the calcium-bound state correlates with T-cell receptor-induced calcium influx.
03:39
In Vivo Calcium Imaging Using Synaptically Localized Calcium Sensors in Drosophila melanogaster
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2025
This video demonstrates a method for in vivo calcium imaging using synaptically localized calcium sensors in flies. It outlines the steps for preparing the flies for microscopy, conducting olfactory conditioning, and visualizing odor-induced calcium influx.
04:46
Calcium Influx Assay to Measure Mitochondrial Calcium Uptake in Cultured Cells
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2025
This video describes a confocal microscopy imaging-based assay to measure mitochondrial calcium uptake in cultured cells. The assay uses the calcium-sensitive fluorescent dye Rhod-2/AM to measure the mitochondrial calcium in permeabilized cultured cells.
06:26
Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
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Cited by 10 •
2011
This protocol describes a method for real-time measurement of mitochondrial calcium fluxes by fluorescent imaging. The method takes advantage of a circularly permutated YFP-based dual-excitation ratiometric calcium sensor (ratiometric pericam-mt) selectively expressed in mitochondria.
FAQs
The acetoxymethyl ester groups increase membrane permeability, allowing Calcium AM to enter cells that would not readily take up the unmodified indicator. After entry, cellular esterases cleave these groups. This processing produces a calcium-sensitive dye that remains inside the cell, enabling fluorescence measurements to reflect intracellular Ca2+ changes during live-cell experiments.
Cellular esterases convert the membrane-permeable form into the retained, calcium-sensitive dye. This two-stage mechanism separates delivery from calcium detection: the esterified indicator enters the cell, while enzymatic cleavage creates the form that responds to Ca2+ binding. Without this intracellular processing, the indicator would not provide the same retained fluorescent readout of calcium dynamics.
Once the dye has been generated inside the cell, its fluorescence changes when it binds Ca2+. Those fluorescence changes provide a readout of alterations in intracellular calcium concentration rather than a direct observation of the underlying cellular event. Interpreting the signal therefore connects optical changes with calcium dynamics associated with cellular activity or signaling.
Researchers can measure Calcium AM responses with fluorescence microscopy, live-cell imaging, or plate-based assays. Microscopy and live-cell imaging support observation of calcium dynamics in individual cells, whereas plate-based formats can assess responses across cell populations. The choice of platform determines whether the experiment emphasizes single-cell behavior or broader population-level measurements.
Calcium AM measurements can characterize cellular activity, signaling pathways, drug responses, and dysfunction associated with disease. Because the indicator reports changes in intracellular calcium, experiments can track how cells respond under different biological or experimental conditions. These readouts help connect calcium dynamics with functional changes observed in individual cells or populations.
In biology research, Calcium AM can monitor calcium dynamics in processes where intracellular Ca2+ acts as an important signal, including muscle contraction, neurotransmission, and cell signaling. Researchers may use fluorescence microscopy, live-cell imaging, or plate-based assays to examine these responses. The resulting measurements support comparisons of cellular activity, pathway behavior, treatment responses, or disease-related dysfunction.