Enzyme-dependent fluorescence recovery of NADH after photobleaching to assess dehydrogenase activity of living tissues
AssigneeThe George Washington University
InventorsMatthew Kay, Angel Moreno, Rafael Jaimes
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Abstract
The invention provides for a system for in vivo real time measurement of NADH recovery kinetics, comprising: 1) a specific pulse sequence to non-destructively, yet effectively, photobleach NADH for measurement of NADH recovery kinetics; 2) illumination light parameters to acquire NADH fluorescence before and after photobleaching, without causing fluorescence bleaching artifacts, for measurement of NADH recovery kinetics; and 3) configurations for devices capable of photobleaching NADH by at least 10% within tissues for effective measurement of NADH recovery kinetics in tissues within a living subject or excised tissues and organs.
Claims34
A catheter to monitor enzyme activity in tissue comprising:
- an ultraviolet illumination device disposed at a distal end of the catheter for exciting cellular, including mitochondrial, NADH of the tissue, wherein the ultraviolet illumination device comprises at least two ultraviolet light-emitting diodes (LEDs) including a first ultraviolet LED to photobleach NADH in the tissue and a second ultraviolet LED to thereafter illuminate the NADH in the tissue for fluorescence imaging;
- a fluorescence sensor at the distal end of the catheter for acquiring a single measurement or an image of NADH fluorescence of the tissue;
- and
- a wire disposed inside a trans-axial conduit within the catheter to connect the fluorescence sensor to a controller located at a proximal end of the catheter,
- wherein a detected fluorescence measurement or image shows a replenishment of fluorescent NADH after photobleaching indicative of cellular dehydrogenase enzyme kinetics within the tissue.
A catheter to monitor enzyme activity in tissue comprising:
- an ultraviolet illumination device disposed at a distal end of the catheter for exciting cellular, including mitochondrial, NADH of the tissue, wherein the ultraviolet illumination device comprises at least two ultraviolet light-emitting diodes (LEDs) including a first ultraviolet LED to photobleach NADH in the tissue and a second ultraviolet LED to thereafter illuminate the NADH in the tissue for fluorescence imaging;
- a fiberscope disposed at the distal end of the catheter for detecting NADH fluorescence from the illuminated tissue at the distal end;
- and
- a fluorescence light detector or camera at a proximal end of the catheter for creating a fluorescence measurement or image from detected NADH fluorescence captured by the fiberscope,
- wherein the fluorescence measurement or image shows replenishment of fluorescent NADH after photobleaching, and this is indicative of cellular dehydrogenase enzyme kinetics within the tissue.
A method for imaging a tissue comprising:
- photobleaching NADH in the tissue, using a first light energy delivered by a first ultraviolet LED;
- illuminating NADH in the tissue, using a second light energy delivered by a second ultraviolet LED;
- and
- imaging fluorescence of illuminated NADH in the tissue to produce a recording of the imaged, illuminated tissue, the recording illustrating cellular dehydrogenase enzyme kinetics in the tissue.
Dependencies are inferred from the claim text.