Fluorophores for super-resolution imaging
The presently-disclosed subject matter includes fluorescent compounds of the following formula: The compounds can be used as probes, dyes, tags, and the like. The presently-disclosed subject matter also includes kits comprising the same as well as methods for using the same to detect a target substance.
1. A compound of the formula:
or a closed form thereof, wherein
(a) X is selected from the group consisting of O, N-alkyl, S, and C(alkyl) 2 ;
Y is —NH 2 or
wherein each Y 1 and Y 2 is each independently selected from the group consisting of H, F, CN, OCH 3 , SO 2 CH 3 , CF 3 , CH 3 , and CO 2 H;
R 1 and R 2 are each independently selected from the group consisting of H, alkyl, and halogen;
R 3 , which can be a substitution at either the 5 position or the 6 position of the ring to which it is bound, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and
R 4 is CH 2 OH;
(b) X is Si(alkyl) 2 ;
Y is —NH 2 ;
R 1 and R 2 are each independently selected from the group consisting of H, alkyl, and halogen;
R 3 , which can be a substitution at either the 5 position or the 6 position of the ring to which it is bound, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, NHS ester, and a targeting moiety; and
R 4 is CH 2 OH;
(c) X is Si(alkyl) 2 ;
Y is
wherein each Y 1 and Y 2 is each independently selected from the group consisting of F, CN, OCH 3 , SO 2 CH 3 , CF 3 , CH 3 , and CO 2 H;
R 1 and R 2 are each independently selected from the group consisting of H, alkyl, and halogen;
R 3 , which can be a substitution at either the 5 position or the 6 position of the ring to which it is bound, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, NHS ester, and a targeting moiety; and
R 4 is CH 2 OH;
(d) X is Si(alkyl) 2 ;
Y is
wherein Y 1 and Y 2 are H;
R 1 and R 2 are each independently selected from the group consisting of alkyl, and halogen;
R 3 , which can be a substitution at either the 5 position or the 6 position of the ring to which it is bound, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, NHS ester, and a targeting moiety; and
R 4 is CH 2 OH; or
(e) X is Si(alkyl) 2 ;
Y is
wherein Y 1 and Y 2 are H;
R 1 and R 2 are H;
R 3 , which can be a substitution at either the 5 position or the 6 position of the ring to which it is bound, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, and a targeting moiety; and
R 4 is CH 2 OH.
2. The compound of claim 1 , according to the following formula:
wherein
Y 1 -Y 4 are each independently selected from the group consisting of H, F, CN, OCH 3 , SO 2 CH 3 , CF 3 , CH 3 , and CO 2 H;
X is selected from the group consisting of O, N-alkyl, S, Si(alkyl) 2 , and C(alkyl) 2 ;
R 1 and R 2 are each independently selected from the group consisting of H, alkyl, and halogen; and
R 3 , which can be a substitution at either the 5 position or the 6 position, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, and a targeting moiety.
3. The compound of claim 1 , according to the following formula:
wherein
R 3 , which can be a substitution at either the 5 position or the 6 position, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety.
4. The compound of claim 1 , according to the following formula:
wherein,
R 3 , which can be a substitution at either the 5 position or the 6 position, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and
Y 1 is H, F, CN, OCH 3 , SO 2 CH 3 , or CF 3 .
5. The compound of claim 1 , according to the following formula:
wherein
R 3 , which can be a substitution at either the 5 position or the 6 position, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and
Y 1 is F, CN, OCH 3 , SO 2 CH 3 , or CF 3 .
6. The compound of claim 1 , according to the following formula:
wherein
R 3 , which can be a substitution at either the 5 position or the 6 position, is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety.
7. The compound of claim 1 , according to a formula selected from the group consisting of:
8. The compound of claim 1 , wherein R 3 is a targeting moiety that is a self-labeling protein tag ligand.
9. The compound of claim 1 , wherein R 3 is a targeting moiety for directing the compound to DNA, microtubules, or lysosomes.
10. The compound of claim 1 , wherein R 3 is a targeting moiety selected from the group consisting of trimethoprim, Taxol, Hoechst, and pepstatin A.
11. A method for detecting a target substance, comprising:
contacting a sample with the compound of claim 1 ; and
detecting an emission light from the compound, the emission light indicating the presence of the target substance.
12. The method of claim 11 , wherein the target substance is selected from a protein, a carbohydrate, a polysaccharide, a glycoprotein, a hormone, a receptor, an antigen, an antibody, a virus, a substrate, a metabolite, an inhibitor, a drug, a nutrient, a growth factor, a lipoprotein, and a combination thereof.
13. The method of claim 11 , wherein the detecting step is performed with a microscope.
14. The method of claim 11 , further comprising a step of exposing the compound to an absorption light that includes a wavelength of about 100 nm to about 1000 nm.
15. The method of claim 11 , wherein the contacting step and the detecting step are performed in a live cell.
16. The method of claim 11 , wherein:
the compound includes a first compound and a second compound;
the first compound being selective for a first target substance and capable of emitting a first emission light;
the second compound being selective for a second target substance and capable of emitting a second emission light, and
the detecting step includes detecting the first emission light that indicates the presence of the first target substance and the second emission light that indicates the presence of the second target substance.
17. The compound of claim 1 , wherein
Y is
wherein Y 1 and Y 2 are selected from the group consisting of
(a) Y 1 is OCH 3 and Y 2 is H,
(b) Y 1 is F and Y 2 is H,
(c) Y 1 is CN and Y 2 is H,
(d) Y 1 is SO 2 CH 3 and Y 2 is H,
(d) Y 1 is CF 3 and Y 2 is H, and
(e) Y 1 and Y 2 are F;
X is Si(alkyl) 2 ;
R 1 and R 2 are H;
R 3 is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and
R 4 is CH 2 OH.
18. The compound of claim 17 , wherein R 3 is a self-labeling protein tag ligand.
19. The compound of claim 17 , wherein R 3 is selected from the group consisting of:
20. The compound of claim 1 , wherein Y is NH 2 ; X is O; R 1 and R 2 are F; R 3 is selected from the group consisting of self-labeling protein tag ligand, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and R 4 is CO 2 H.
21. The compound of claim 20 , wherein R 3 is a self-labeling protein tag ligand.
22. The compound of claim 20 , wherein R 3 is selected from the group consisting of:
23. The compound of claim 1 , wherein
Y is
wherein Y 1 and Y 2 are selected from the group consisting of
(a) Y 1 is OCH 3 and Y 2 is H,
(b) Y 1 is F and Y 2 is H,
(c) Y 1 is CN and Y 2 is H,
(d) Y 1 is SO 2 CH 3 and Y 2 is H,
(d) Y 1 is CF 3 and Y 2 is H, and
(e) Y 1 and Y 2 are F;
X is O, N-alkyl, S, and C(alkyl) 2 ;
R 1 and R 2 are H, alkyl, or halogen;
R 3 is selected from the group consisting of H, self-labeling protein tag ligand, CO 2 H, CO 2 CH 3 , CO 2 t-Bu, N-hydroxysuccinimidyl (NHS) ester, and a targeting moiety; and
R 4 is CH 2 OH.
24. The compound of claim 23 , wherein R 3 is selected from the group consisting of: