Fluorescent membrane intercalating probes and methods for their use
The invention relates to a family of dyes which fluoresce in the UV-VIS, far red and near infrared wavelengths of the spectrum and possess asymmetric lipophilic alkyl chains. The dyes of the invention are soluble in commercially available membrane staining dyes, are useful as probes for rapidly staining lipophilic structures such as membranes in cells or isolated from cells, and are well-retained therein. Methods of utilizing the dyes to detect stained cells both in vivo and in vitro are also disclosed.
1. A cyanine dye having the following chemical structure:
wherein Z is a biologically compatible counterion.
2. The cyanine dye of claim 1 , wherein the counterion is selected from the group consisting of chloride, bromide, iodide, sulfate, alkanesulfonate, arylsulfonate, phosphate, perchlorate, tetrafluoroborate, nitrate, an anion of an aromatic carboxylic acid and an anion of an aliphatic carboxylic acid.
3. The cyanine dye of claim 1 , wherein the counterion is iodide.
4. An in vivo method for detecting abnormal cell shedding rates of mature surface epithelial cells of a warm-blooded animal, comprising the steps of:
labeling mature surface epithelial cells at a target site of the animal with the cyanine dye of claim 1 , and
monitoring the site for the presence or absence of the cyanine dye following said labeling step.
5. The method of claim 4 , wherein the site is a mucosal surface.
6. The method of claim 4 , wherein the mucosal surface lines the surface of the gastrointestinal tract, the respiratory tract, or the genitourinary tract.
7. The method of claim 5 wherein the site is a mucosal surface of the stomach.
8. The method of claim 5 wherein the site is a mucosal surface of the colon.
9. The method of claim 4 , wherein cell shedding is detected by observing changes in the level of the cyanine dye at the site at a pre-selected time following said labeling step.
10. The method of claim 4 , further comprising the step of exciting the target site with a light source wherein the light source has a wavelength of from about 600 nm to 1000 nm.
11. The method of claim 4 , wherein the epithelial cells are labeled by direct application of a labeling composition comprising the cyanine dye to the site.
12. The method of claim 4 , wherein said labeling step comprises applying to mature surface epithelial cells at a target side a labeling composition comprising the cyanine dye.
13. The method of claim 12 , wherein said monitoring step comprises observing the rate at which the cyanine dye is lost from the target site.
14. The method of claim 12 , wherein said labeling composition further comprises a pharmaceutically acceptable vehicle.
15. The method of claim 11 , wherein said labeling composition further comprises a pharmaceutically acceptable vehicle.
16. The method of claim 4 , wherein the counterion is selected from the group consisting of chloride, bromide, iodide, sulfate, alkanesulfonate, arylsulfonate, phosphate, perchlorate, tetrafluoroborate, nitrate, an anion of an aromatic carboxylic acid and an anion of an aliphatic carboxylic acid.
17. The method of claim 4 , wherein the counterion is iodide.