ENERGY METABOLISM-DEPENDENT CONTRAST REAGENT IN NEAR INFRARED REGION

本发明的目的在于提供一种测定细胞或组织的能量状态的方法，特别是提供一种具有高透过率、能够测定器官等厚组织的能量状态的测定方法。其特异性地掺入细胞线粒体中并显示不同的吸收光谱或荧光强度;通过测量分别选自730-850 nm波长范围和730-900 nm波长范围的两个不同波长处的吸光度之间的差异，可以适当地检测吸光度的变化，优选730 nm和800 nm ;掺入线粒体的罗丹明800在线粒体内沿着线粒体膜电位的变化形成复合物，并提供不同的荧光光谱。 ...更多信息 因此，本研究提供了以下内容。(1)一种用于确定细胞或组织的能量状态的方法，其包括使罗丹明800与细胞和/或组织接触，并测量分别选自730-850 nm的波长区域和730-900 nm的波长区域，优选730 nm和800 nm的波长区域的两个不同波长下的罗丹明800之间的差。(2)一种用于确定细胞或组织的能量状态的方法，其包括使罗丹明800与细胞和/或组织接触并测量由线粒体内形成的复合物引起的荧光强度。(3)上述（2）的方法，其特征在于，还包括在选自600-680 nm的范围的波长下进行荧光激发，并在选自680-730 nm的范围且与激发波长不同的波长下测定荧光发射强度，特别是在660 nm下进行荧光激发，并在692 nm下测定荧光发射强度。(4)一种通过上述（1）~（3）的方法确定肝细胞和/或心肌细胞的能量状态的方法。(5)上述（1）~（4）的方法，其中，细胞的能量状态是指不可逆的细胞死亡（膜电位的损失）。少

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