ENERGY METABOLISM-DEPENDENT CONTRAST REAGENT IN NEAR INFRARED REGION

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It is an object of the present research to provide a method for determining the energy state of cell or tissue, more particularly, a method for determining energy state, which has a high transmittivity and therefore is capable of determining a thick tissue such as an organ.As a results of intensive studies, there have been found that energy state of cell or tissue can be determined by bringing Rhodamine 800, which is specifically incorporated into cell mitochondria and shows varying absorption spectrum or fluorescence intensity ; that the changes in the absorbance can be appropriately detected by measuring the difference between absorbances at two different wavelengths respectively selected from a wavelength range of 730-850 nm and a wavelength range of 730-900 nm, preferably 730 nm and 800 nm ; and that Rhodamine 800 incorporated into mitochondria forms a complex within the mitochondria along the changes in mitochondrial membrane potential and affords different fluorescence spectra.Ac … More cordingly, the present research provides the following.(1) A method for determining an energy state of a cell or tissue, which comprises bringing Rhodamine 800 into contact with the cell and/or the tissue and measuring the difference between absorbances at two different wavelengths respectively selected from a wavelength region of 730-850 nm and a wavelength region of 730-900 nm, preferably 730 nm and 800 nm.(2) A method for determining an energy state of a cell or tissue, which comprises bringing Rhodamine 800 into contact with the cell and/or the tissue and measuring fluorescence intensity caused by a complec formed within mitochondria.(3) A method for (2) above, which further comprises fluorescent excitation at a wavelength selected from the range of 600-680 nm and measuring a fluorescence emission intensity at a wavelength selected from the range of 680-730 nm and different from the excitation wavelength, particularly, fluorescent excitation at 660 nm and measuring a fluorescence emission intensity at 692 nm.(4) A method for determining an energy state of hepatocytes and/or myocardial cells by the method of (1) to (3) above.(5) The method of (1) to (4) above, wherein the energy state of cell means an irreversible cell death (loss of membrane potential). Less