Fluo-3

Fluo-3 is a fluorescence indicator of intracellular calcium (Ca2+), developed by Roger Y. Tsien.[1] It is used to measure Ca2+ inside living cells in flow cytometry, and confocal laser scanning microscopy using visible light excitation (compatible with argon laser sources operating at 488 nm). Fluo-3 and derivatives (Fluo-4, Fluo-5 etc) have also been widely used with two-photon excitation microscopy. Fluo-3 is an essentially nonfluorescent compound, but upon binding of Ca2+ its fluorescence increases sharply with an emission maximum at 525 nm suitable for conventionally used detectors designed for fluorescein isothiocyanate (FITC) measurements. This large change in fluorescence coupled with a good yield of photons provides very high contrast which allowed the detection of microscopic Ca2+ release events inside cells called "Calcium sparks".[2] Whereas the salts of fluo-3 are unable to penetrate cells, loading can be achieved using its acetoxymethyl (AM) ester derivative. Once inside the cell, unspecific esterases cleave the ester effectively trapping fluo-3.[3]

Fluo-3
Names
Preferred IUPAC name
2,2′-{[2-(2-{2-[Bis(carboxymethyl)amino]-5-(2,7-dichloro-6-hydroxy-3-oxo-3H-xanthen-9-yl)phenoxy}ethoxy)-4-methylphenyl]azanediyl}diacetic acid
Identifiers
3D model (JSmol)
ChEMBL
ChemSpider
UNII
  • InChI=1S/C36H30Cl2N2O13/c1-18-2-4-24(39(14-32(43)44)15-33(45)46)30(8-18)51-6-7-52-31-9-19(3-5-25(31)40(16-34(47)48)17-35(49)50)36-20-10-22(37)26(41)12-28(20)53-29-13-27(42)23(38)11-21(29)36/h2-5,8-13,41H,6-7,14-17H2,1H3,(H,43,44)(H,45,46)(H,47,48)(H,49,50)
    Key: OZLGRUXZXMRXGP-UHFFFAOYSA-N
  • InChI=1/C36H30Cl2N2O13/c1-18-2-4-24(39(14-32(43)44)15-33(45)46)30(8-18)51-6-7-52-31-9-19(3-5-25(31)40(16-34(47)48)17-35(49)50)36-20-10-22(37)26(41)12-28(20)53-29-13-27(42)23(38)11-21(29)36/h2-5,8-13,41H,6-7,14-17H2,1H3,(H,43,44)(H,45,46)(H,47,48)(H,49,50)
    Key: OZLGRUXZXMRXGP-UHFFFAOYAI
  • O=C(O)CN(c5ccc(cc5OCCOc4c(N(CC(=O)O)CC(=O)O)ccc(C=1c3c(OC=2C=1\C=C(\Cl)C(=O)C=2)cc(O)c(Cl)c3)c4)C)CC(=O)O
Properties
C36H30Cl2N2O13
Molar mass 769.54 g·mol−1
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
Infobox references

As calcium is a key second messenger within cells, the specific properties of fluo-3 enable researchers to investigate the time-resolved dynamics of intracellular signal transduction in a diverse range of cells.[4][5]

References
  1. Tsien, R.Y. (1980). "New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures". Biochemistry. 19 (11): 2396–2404. doi:10.1021/bi00552a018.
  2. Cheng, H.; Lederer, W.J.; Cannell, M.B. (1993). "Calcium Sparks - Elementary Events Underlying Excitation-Contraction Coupling in Heart-Muscle". Science. 262 (5134): 740–744. Bibcode:1993Sci...262..740C. doi:10.1126/science.8235594. PMID 8235594.
  3. Haugland, RP. Handbook of Fluorescent Probes and Research Products. Molecular Probes, 2010
  4. Gamsjäger, T. Flow Cytometry of Intracellular Calcium in Platelets. Grin, 2012
  5. Lambert, DG. Calcium Signaling Protocols. Humana Press, 2006
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