Beliaeva T N, Krolenko S A, Leont'eva E A, Mozhenok T P, Salova A V, Faddeeva M D
Tsitologiia. 2009;51(2):103-10.
Using spectral scanning regime of Leica TCS SL confocal microscope, acridine orange (AO) fluorescence spectra in nuclei and cytoplasms of living myoblasts L6J1 and frog single muscle fibres have been studied. AO fluorescence spectra in salt solutions dependent on free AO concentrations and in AO complexes with DNA have also been obtained for comparison. Myoblasts nuclei fluoresced in green spectral region with maximum at approximately 530 nm (corresponding AO monomers fluorescence), nucleoli fluoresced most brightly. Nuclear chromatin fluoresced not uniformly in these cells. We saw similar to myoblasts AO emission in nucleoli and nuclei of frog single muscle fibres. The uniformed weak green fluorescence was observed for myoblast cytoplasm. As to the muscle fibres sarcoplasm, we saw also AO green fluorescence in A-discs. In myoblasts and muscle fibre cytoplasm we saw the fluorescent red, yellow and green granules which were acidic organelles. The comparison of AO fluorescence spectra in living cells with fluorescence spectra of different AO concentrations and complexes of AO with DNA in buffer solutions allows estimation of AO concentration in acidic granules which is of interest in the investigation of cellular organelles functions in the processes of intracellular transport, adaptation, apoptosis and a number of pathological conditions.
利用徕卡TCS SL共聚焦显微镜的光谱扫描模式,研究了活的成肌细胞L6J1和青蛙单根肌纤维细胞核与细胞质中吖啶橙(AO)的荧光光谱。还获得了盐溶液中依赖于游离AO浓度的AO荧光光谱以及AO与DNA复合物的荧光光谱用于比较。成肌细胞核在绿色光谱区域发出荧光,最大值约为530 nm(对应AO单体荧光),核仁发出的荧光最亮。这些细胞中的核染色质荧光不均匀。我们在青蛙单根肌纤维的核仁和细胞核中观察到了与成肌细胞类似的AO发射。成肌细胞细胞质观察到均匀的弱绿色荧光。至于肌纤维肌浆,我们在A带中也看到了AO绿色荧光。在成肌细胞和肌纤维细胞质中,我们看到了呈荧光的红色、黄色和绿色颗粒,它们是酸性细胞器。将活细胞中AO的荧光光谱与缓冲溶液中不同AO浓度以及AO与DNA复合物的荧光光谱进行比较,可以估算酸性颗粒中的AO浓度,这对于研究细胞器在细胞内运输、适应、凋亡以及许多病理状况过程中的功能具有重要意义。