Fluo-5n Am
Fluo-5n Am - Related Videos
08:05
Staining the Cytoplasmic Ca2+ with Fluo-4/AM in Apple Pulp
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Cited by 1 •
2021
Isolated protoplasts of apple pulp cells were loaded with a calcium fluorescent reagent to detect cytoplasmic Ca2+ concentration.
03:13
Mitochondrial Calcium Uptake Assay: A Plate Reader-Based Method for Measuring Calcium Uptake in Isolated Mitochondria
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2025
This video describes a plate reader-based assay to measure mitochondrial calcium uptake in isolated mitochondria. The assay analyzes the fluorescence signal of a calcium-sensitive fluorescent dye to study the kinetics of mitochondrial calcium uptake and calcium overload.
![Development and Characterization of <em>In Vitro</em> Microvessel Network and Quantitative Measurements of Endothelial [Ca<sup>2+</sup>]<sub>i </sub>and Nitric Oxide Production](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F54014.jpg%3Fv%3D2026-09-01%252006%253A38%253A51&w=750&q=75)
09:39
Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production
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Cited by 7 •
2016
Primary human umbilical vein endothelial cells (HUVECs) were grown to confluence within a microfluidic network device. The endothelial cell junction and F-actin distributions were illustrated and the changes in intracellular calcium concentration and nitric oxide production in response to adenosine triphosphate (ATP) were quantified in real-time at individual cell levels.
08:43
Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
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Cited by 28 •
2017
Mitochondria can utilize the electrochemical potential across their inner membrane (ΔΨm) to sequester calcium (Ca2+), allowing them to shape cytosolic Ca2+ signaling within the cell. We describe a method for simultaneously measuring mitochondria Ca2+ uptake and ΔΨm in live cells using fluorescent dyes and confocal microscopy.
07:15
Single-Cell Characterization of Calcium Influx and HIV-1 Infection using a Multiparameter Optofluidic Platform
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Cited by 1 •
2021
Here, we present a protocol in which single cells are monitored for acute events and productive HIV-1 infection on a nanofluidic device. Imaging data define virus-host receptor interactions and signaling pathway dynamics. This is the first method for nanofluidic high-throughput longitudinal single-cell culture and imaging to study signaling kinetics and molecular interactions.
FAQs
Its low calcium affinity helps the indicator remain informative in compartments where calcium concentrations are relatively high. A higher-affinity probe may respond strongly under those conditions and provide less useful discrimination across changing levels. Fluo-5N Am therefore supports measurements of calcium loading, release, and redistribution in calcium-rich intracellular environments such as the endoplasmic or sarcoplasmic reticulum.
The acetoxymethyl ester temporarily masks properties that would otherwise limit membrane passage, allowing Fluo-5N Am to enter cells. Once inside, intracellular esterases cleave the ester groups and convert the dye into its fluorescent, calcium-sensitive form. This enzymatic processing also helps retain the active indicator within the cell for monitoring intracellular calcium dynamics.
Calcium binding changes the fluorescence behavior of Fluo-5N Am. By tracking fluorescence with microscopy or a related assay, researchers can follow changes in intracellular calcium over time rather than observing calcium directly. The resulting signal can reveal whether calcium is being loaded into a compartment, released from it, or redistributed within the cell.
Fluo-5N Am can report changing fluorescence as calcium moves between intracellular locations or changes in availability within a compartment. This makes the probe useful for examining dynamic processes, including calcium loading followed by release and subsequent redistribution. In biology, those measurements help connect organelle calcium behavior with cellular signaling and maintenance of calcium homeostasis.
A typical workflow introduces the membrane-permeant dye to cells, allows intracellular esterases to cleave its AM ester groups, and then measures fluorescence with microscopy or a related assay. The experiment is designed so that the processed indicator remains inside cells while calcium-dependent fluorescence changes are recorded during loading, release, or redistribution events.
The probe supports studies of intracellular calcium dynamics, especially in organelles with relatively high calcium levels. Researchers can use fluorescence measurements to examine calcium storage and release in the endoplasmic or sarcoplasmic reticulum, relate those changes to cellular signaling, and assess how calcium redistribution contributes to broader regulation of cellular homeostasis.