Fluo-3 Probes

Fluo-3 is a fluorescent indicator of intracellular calcium (Ca²⁺). It is used to measure Ca²⁺ in living cells using visible light excitation (compatible with an argon laser source operating at 488 nm) in flow cytometry and confocal laser scanning microscopy. Fluo-3 is a basic non-fluorescent compound, but when combined with Ca²⁺, its fluorescence will increase sharply and its maximum emission will occur at 525 nm. It is suitable for the design of fluorescein isothiocyanate (FITC) measurement. Routine detector. This large change in fluorescence, coupled with good photon yield, provides very high contrast so that tiny Ca²⁺ release events inside cells called "calcium sparks" can be detected. Although the salt of fluo-3 cannot penetrate cells, it can be loaded using its acetoxymethyl (AM) ester derivative. Once in the cell, non-specific esterases cleave esters, effectively capturing fluo-3. Because calcium is a key secondary messenger within cells, the special properties of fluo-3 allow researchers to study the time-resolved kinetics of intracellular signal transduction in various cells.

Figure 1. Structure of Fluo-3 Probes.

Introductions

Fluo 3 is almost non-fluorescent if it exists as a free ligand, but when it is combined with calcium ion Ca²⁺, its fluorescence will increase 60 to 80 times. It is currently the most commonly used calcium ion fluorescent probe. The laser confocal fluorescence microscope has an argon laser, so Fluo 3 can be widely used in this microscope. This long wave from the fluorescent signal also facilitates reducing light damage to the sample cells. Fluo 3 can also be used to detect cleavable chelated calcium or other forms of calcium under UV light. Fluo 3-AM is an acetomethyl derivative of Fluo 3, which can easily enter cells through culture. Fluo 3-AM (Calcium Fluorescent Probe) needs to be formulated with anhydrous DMSO. Fluo 3-AM is a fluorescent dye that penetrates cell membranes. After entering the cell, Fluo 3-AM can be cleaved by intracellular esterase to form Fluo 3, which is then retained in the cell. Fluo 3 can be combined with calcium ions. Combining calcium ions can produce strong fluorescence, with a maximum excitation wavelength of 506 nm and a maximum emission wavelength of 526 nm.

References:

Latt, SA.; et al. Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence. Journal of Histochemistry and Cytochemistry. 1975, 23 (7): 493–505.
Cheng, H.; et al. Calcium Sparks - Elementary Events Underlying Excitation-Contraction Coupling in Heart-Muscle. Science. 1993, 262 (5134): 740–744.
Haugland, RP. Handbook of Fluorescent Probes and Research Products. Molecular Probes, 2010

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