In vivo visualization of lymphatic tissue
View Patent ↗Aspects of the invention include methods for visualizing lymphatic tissue in an ocular region, e.g., the cornea, of a living subject, such as a mouse or human. In certain embodiments, the methods include contacting the region with a fluorescently-labeled lymphatic tissue-specific dye, e.g., FITC or rhodamine labeled dextran, and detecting the labeled dye to visualize lymphatic tissue in the region, where the visualization may vary, from an image obtained at a single time to a video over a period of time. The invention finds use in a variety of different applications, including research and therapeutic applications.
1. A method for visualizing lymphatic tissue in an ocular region of a living subject, the method comprising:
contacting the ocular region with a detectably-labeled lymphatic tissue-specific dye via injection into a subconjunctival space; and
detecting the labeled dye to visualize lymphatic tissue in the ocular region of the living subject;
wherein the detectably-labeled lymphatic tissue-specific dye comprises a fluorescent label covalently bound to a physiologically-acceptable polysaccharide.
2. The method according to claim 1 , wherein the ocular region is the cornea.
3. The method according to claim 1 , wherein the method comprises inducing lymphangiogenesis in the ocular region.
4. The method according to claim 3 , wherein the ocular region is the cornea and lymphangiogenesis is induced in the cornea using a suture protocol, an implantation protocol or a transplantation protocol.
5. The method according to claim 1 , wherein the polysaccharide has a molecular weight ranging from 1,000 kDa up to 10,000 kDa.
6. The method according to claim 5 , wherein the polysaccharide is dextran.
7. The method according to claim 1 , wherein the detecting comprises obtaining an image at a single time.
8. The method according to claim 1 , wherein the detecting comprises obtaining two or more images at different times.
9. The method according to claim 1 , wherein the detecting comprises obtaining a video over a period of time.
10. The method according to claim 1 , wherein the method further comprises visualizing blood vessels in the ocular region.
11. The method according to claim 1 , wherein the method further comprises contacting the ocular region with a candidate agent and evaluating the impact of the candidate agent on at least one of lymphatic and blood vessel activity in the ocular region.
12. The method according to claim 11 , wherein the candidate agent is covalently bound to the detectably-labeled lymphatic tissue-specific dye.
13. The method according to claim 1 , wherein the method further comprises contacting the ocular region with a therapeutic agent conjugated to the detectably-labeled lymphatic tissue-specific dye.
14. The method according to claim 1 , wherein the method further comprises contacting the ocular region with a detectably-labeled lymphatic tissue-specific antibody and detecting the antibody.
15. The method according to claim 14 , wherein the detectably-labeled lymphatic tissue-specific dye comprises a fluorescent label having a first maximum emission wavelength, and the detectably-labeled lymphatic tissue-specific antibody comprises a fluorescent label having a second maximum emission wavelength that is different from the first maximum emission wavelength.
16. The method according to claim 1 , wherein the living subject is a mammal.
17. The method according to claim 1 , wherein the polysaccharide is selected from a pullulan, a dextrin, a dextran sulfate, and a dextran.
18. The method according to claim 17 , wherein the fluorescent label comprises a physiologically-acceptable xanthene dye.
19. The method according to claim 18 , wherein the fluorescent label is selected from a fluorescein dye and a rhodamine dye.
20. The method according to claim 19 , wherein the polysaccharide is dextran.
21. The method according to claim 1 , wherein the method for visualizing lymphatic tissue consists essentially of:
contacting the cornea with a detectably-labeled lymphatic tissue-specific dye that comprises a fluorescent label covalently bound to a physiologically-acceptable polysaccharide having a molecular weight ranging from 1,000 kDa up to 10,000 kDa; and
detecting the labeled dye to visualize lymphatic tissue in the cornea of the living subject.
22. The method according to claim 21 , wherein the polysaccharide is selected from a pullulan, a dextrin, a dextran sulfate and a dextran.
23. The method according to claim 22 , wherein the polysaccharide is dextran.