Table of Contents
How do you measure calcium concentration in cells?
Changes in intracellular calcium concentration can be measured using the calcium sensitive fluorescent ratiometric dye fura-2 AM. This method is a high throughput way to measure agonist mediated calcium responses.
How does fluo 4 work?
Fluo-4 is an analog of Fluo-3 with the two chlorine substituents replaced by fluorines, which results in increased fluorescence excitation at 488 nm that gives higher fluorescence signal. Fluo-4 has its absorption maximum at 494 nm, thus making it excitable by the argon-ion laser.
Which filter cube did you use to visualize fluo 4?
Fluo-4-AM emission was captured using GaAsP high QE 32 channel spectral array detectors and a standard green fluorescence filter cube. In parallel, Hoechst 33342 blue fluorescence was captured from the same fields to visualize nuclei within the imaging plane.
How is cytosolic calcium measured?
Detecting changes in cytosolic calcium is normally performed using a live cell imaging set up with calcium binding dyes that exhibit either an increase in fluorescence intensity or a shift in the emission wavelength upon calcium binding.
How is calcium influx measured?
There are several directly or indirectly ways to measure intracellular Ca2+ influx:
- Using Ca2+ sensitive dye such as FURa-2 as recommended to you by others.
- using intracellular Ca2+ sensitive microelectrode.
- Performing Whole cell patch clamp recording.
What is Kd of calcium?
Ca2+ affinities are relatively high (Kd ~170-185 nM).
What is the role of calcium in neurons?
In neurons calcium plays a dual role as a charge carrier and an intracellular messenger. Calcium signals regulate various developmental processes and have a key role in apoptosis, neurotransmitter release and membrane excitability.
What is a calcium flux?
Indo-1 is the ratio metric calcium indicator dye most commonly used in flow cytometry because of its shift in emission frequency when excited at a single wavelength. …