Fluorogenic probe using a Mislow-Evans rearrangement for real-time imaging of hydrogen peroxide

1. A compound having the structure:

wherein:

R 1 is a fluorescent dye moiety that does not fluoresce when incorporated in the compound or fluoresces with a different intensity or peak wavelength when incorporated in the compound; and

R 2 is optionally substituted and is alkyl, heteroalkyl, aryl, heteroaryl, or any combination of the proceeding.

2. The compound of claim 1 , wherein R 1 is a xanthene dye moiety, and R 2 is phenyl, optionally substituted aryl, optionally substituted heteroaryl, or C 1 -C 10 alkyl.

3. The compound of claim 2 , wherein R 1 is

4. The compound of claim 2 , wherein R 1 is

5. The compound of claim 2 , wherein R 2 is phenyl.

6. The compound of claim 2 , wherein R 2 is a nitrogen-substituted heteroaryl moiety.

7. The compound of claim 6 , wherein R 2 is a pyridine moiety.

8. The compound of claim 6 , wherein R 2 is

9. The compound of claim 1 having the structure:

10. The compound of claim 1 having the structure:

11. A method of identifying or visualizing the presence of H 2 O 2 in a cell, tissue, organ, or organism, comprising contacting the cell, tissue, organ, or organism with a compound according to claim 1 , illuminating the cell, tissue, organ, or organism with light including or at an excitation wavelength for the reaction product of the compound with H 2 O 2 , and detecting fluorescent emission from the reaction product.

12. The method of claim 11 , further comprising obtaining an image of the cell, tissue, organ, or organism at or including light at the excitation wavelength for the reaction product of the compound with H 2 O 2 , optionally using fluorescence microscopy.

13. The method of claim 11 , further comprising obtaining a spectrograph of light emitted by the reaction product, and optionally quantifying H 2 O 2 based on light emitted by the reaction product.

14. The method of claim 11 , further comprising sorting cells using a fluorescence-activated cell sorter, based on an amount of light emitted by the reaction product in each cell.

15. The method of claim 14 , further comprising classifying cells sorted based on the amount of light emitted by the reaction product in each cell, and quantifying H 2 O 2 production in the sorted cells based on the number of cells sorted.

16. A method of identifying or quantifying the presence of H 2 O 2 in a sample, comprising adding to or mixing in the sample a compound according to claim 1 , illuminating the sample with light including or at an excitation wavelength for the reaction product of the compound with H 2 O 2 , and detecting fluorescent emission from the reaction product.

17. The method of claim 16 , wherein the sample is contained within a cuvette, multi-well plate, or lateral flow device, optionally wherein the cuvette, multi-well plate, or lateral flow device are disposable, or form part of a disposable article.

18. The method of claim 16 , wherein the sample is a biological sample, such as blood, plasma, serum, urine, cerebrospinal fluid, mucus, lymph, cell lysate or a fraction or derivative thereof, or conditioned cell culture medium.

19. The method of claim 16 , wherein the H 2 O 2 is converted to a different compound in an enzyme-linked immunoassay, e.g., using catalase to decompose the H 2 O 2 to water and oxygen, and the identification or quantification of the presence of the H 2 O 2 measures the presence of or quantity of an analyte or binding activity detected by the immunoassay.