Comparison of techniques to determine the subcellular localization of photosensitizers / 中华物理医学与康复杂志

ABSTRACT

Objective To compare and evaluate Organelle-cell fluorescence intensity ratio analysis established in this study with other techniques to determine the subcellular localization of photosensitizers. Methods CCD fluorescence microscopy imaging system was applied and a kind of special organelle probe BODIPY was selected to label Golgi body. Directly observing, pseudo-color fusing, wave-shape comparing, correlation coefficient calcula-ting and Organelle-cell fluorescence intensity ratio analysis were adopted, respectively, to study the intracellular distribution of domestic photosensitizer Hematoporphyrin monomethyl ether (HMME). Results Fluorescence distributing modes of HMME and BODIPY were similar with each other. There ware yellow space-overlap areas in the fusion image. Wave body of changing curve of gray scale value of all pixels in the straight line in two images corresponding to space coordinate was similar with each other. Correlation coefficient in each pixel between fluorescence intensity of HMME and that of BODIPY was 0.602 4. With increasing of parameter m, namely degree of organelles congregated reducing, the average fluorescence intensity ratio (J_1/J_2) of Golgi complex was decreasing, the two parameters po-ssessed obvious relativity. P