Two-photon probe for real-time monitoring of intracellular magnesium ions, method for preparing the two-photon probe and method for real-time monitoring of intracellular magnesium ions using the two-photon probe
View Patent ↗A two-photon probe for real-time monitoring of intracellular magnesium ions is provided. Specifically, the two-photon probe is represented by Formula 1: wherein R is H or CH 2 OCOCH 3 . The two-photon probe is very suitable for real-time imaging of intracellular magnesium ions. The two-photon probe shows 17-fold two-photon excited fluorescence enhancement in response to Mg 2+ , which is 7-fold stronger than commercial probes, thus enabling staining of cells in a greatly reduced amount. In addition, the two-photon probe has a sufficiently low molecular weight to stain cells and is very suitable for monitoring Mg 2+ ions present in the deep tissue. Furthermore, the two-photon probe can be effectively used for the quantitative as well as qualitative detection of intracellular magnesium ions. Further provided are a method for preparing the two-photon probe and a method for real-time monitoring of intracellular magnesium ions using the two-photon probe.
1. A two-photon probe for real-time monitoring of intracellular magnesium ions, represented by Formula 1:
wherein R is H or CH 2 OCOCH 3 .
2. A method for preparing a two-photon probe for real-time monitoring of intracellular magnesium ions, represented by Formula 1:
wherein R is H or CH 2 OCOCH 3 ,
the method comprising reacting 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and the compounds of Formulae 2 and 3:
3. The method according to claim 2 , wherein the compound of Formula 2 is prepared by reacting methyl bromoacetate, Na 2 HPO 4 , NaI and the compound of Formula 4:
4. The method according to claim 3 , wherein the compound of Formula 4 is prepared by reacting CH 3 NH 2 .HCl, Na 2 S 2 O 3 , NaOH, H 2 O and the compound of Formula 5:
5. The method according to claim 4 , wherein the compound of Formula 5 is prepared by reacting HBr with the compound of Formula 6:
6. A method for real-time monitoring of intracellular magnesium ions, the method comprising introducing the two-photon probe according to claim 1 into cells and imaging two-photon excited fluorescence emitted from the two-photon probe.
7. The method according to claim 6 , wherein the two-photon excited fluorescence images are obtained in the wavelength range of 500 to 620 nm.
8. The method according to claim 6 , wherein the intracellular magnesium ion concentration is quantitatively determined by Equation 1:
[Mg 2+ ]=K d [( F−F min )/( F max −F )] (1)
where K d is the dissociation constant of the two-photon probe for Mg 2+ , F is the observed two-photon fluorescence intensity, F min is the minimum fluorescence intensity, and F max is the maximum fluorescence intensity.