Bistrifilate-based fluorogenic probes for detection of superoxide anion radical
The invention provides fluorogenic compounds and probes that can be used as reagents for measuring, detecting and/or screening superoxide. The fluorogenic compounds of the invention can produce fluorescence colors, such as green, yellow, red, or far-red. The invention further provides fluorogenic compounds for selectively staining superoxide in the mitochondria of living cells. The invention also provides methods that can be used to measure, directly or indirectly, the presence and/or amount of superoxide in chemical samples and biological samples such as cells and tissues in living organisms, and a high-throughput screening methods for detecting or screening superoxide or compounds that can increase or decrease the level of superoxide in chemical and biological samples.
1. A fluorogenic probe composition for detecting superoxide, comprising a compound having one of the following formulas 3-4, 9, 10 and 13-20:
wherein the compound reacts with superoxide to form one or more fluorescent compounds.
2. The fluorogenic probe composition of claim 1 , further comprising a carrier.
3. The fluorogenic probe composition of claim 1 , wherein the fluorogenic probe composition further comprises a solvent, an acid, a base, a buffer solution, or a combination thereof.
4. A method for detecting the presence of, and/or determining the level of superoxide in a sample, comprising:
a) contacting the fluorogenic probe composition of claim 1 with the sample to form a fluorescent compound; and
b) determining fluorescence property of the fluorescent compound.
5. The method of claim 4 , wherein the sample is a chemical sample or biological sample.
6. The method of claim 4 , wherein the sample is a biological sample comprising a microorganism, or a cell or tissue.
7. A method for detecting the presence of, or determining the level of superoxide in vivo in an organism, comprising:
a) administering the fluorogenic probe composition of claim 1 to the organism to form a fluorescent compound; and
b) determining fluorescence property of the fluorescent compound.
8. A high-throughput screening method for detecting the presence of, or determining the level of, superoxide in samples, wherein the high-throughput method comprises the steps of:
a) contacting the fluorogenic probe composition of claim 1 with the samples to form one or more fluorescent compounds; and
b) determining fluorescence properties of the fluorescent compounds to determine the presence and/or amount of peroxynitrite in the samples.
9. A high-throughput method for screening one or more target compounds that increase or decrease the level of superoxide, wherein the high-throughput method comprises the steps of:
a) contacting the fluorogenic probe composition of claim 1 with target compounds to form one or more fluorescent compounds; and
b) measuring fluorescence properties of the florescent compounds to determine the presence and/or amount of the target compounds.