游离锌离子形态学检测方法探究_医学论文.docVIP

游离锌离子形态学检测方法探究_医学论文 游离锌离子形态学检测方法探究_医学论文 【摘要】 目的:使用ivZnSAMG、ZnSeAMG、iZnSAMG、TSQ荧光、Zinquin荧光对海马苔藓游离锌离子进行染色和比较。方法: ivZnSAMG：采用Na2S溶液灌流的锌离子结合方式；ZnSeAMG：采用事先注射硒酸钠的体内锌离子结合方式；iZnSAMG：采用取动物新鲜组织浸泡在浸染液中的锌离子结合方式；以上三种方法处理后均采用相同的AMG孵育显色。TSQ、Zinquin荧光 :均为新鲜组织取材，液氮处理后荧光下观察。结果：在小鼠海马，所有染色方法均能标记出苔藓纤维的染色，各种染色在苔藓纤维标记有细节上的差异。所有染色方法中特异性最高的是ZnSeAMG；但其敏感性相对较差。而ivZnSAMG的敏感性最高，而其特异性则相对较差。结论：本实验中使用的游离锌离子染色方法都能成功的对游离锌离子进行标记，而这些染色方法的特异性和敏感性各有不同，对染色细节的雕琢能力也各有不同，因此可以根据这些方法的各自特点在实验选取适合的方法。 【关键词】 游离锌离子；N荧光技术；金属自显影；Zinquin荧光技术；苔藓纤维 ＡｂｓｔｒａｃｔObjective:To compare the difference dying results of the free zinc in the hippocampus mossy fiber between the different methods such as Na2SAMG、ZnSeAMG、iZnSAMG、TSQ fluorescence and Zinquin fluorescence. Method: Na2SAMG：zinc was combined with sodium sulphide perfusion ZnSeAMG：zinc was combined with injection sodium selenite in advanced；iZnSAMG：the animals fresh tissue immersed in the Timms immersion fluid. Aboved three methods were followed by the same AMG method. TSQ、Zinquin fluorescence :the animals fresh tissues were observed under the fluoresence microscope after the frozen in the liquid nitrogen. Results: In the hippocampus of the mice, the mossy fiber could be stained by all of the aboved methods. However there were some differences between them in details. Among all of the methods mentioned above, the ZnSeAMG method had the most specificity and the lowest sensibility but the ivZnSAMG method on the contrary properly. Conclusion:All of the above methods could catch the zinc, but the specificity and sensibility are different respectively. Therefore these different methods are used in the different experiments with the different destination. Ｋｅｙ　ｗｏｒｄｓ free zinc N(6methoxy8quinolyl)ptoluenesulfonamide (TSQ) fluorescence AMG (autometallography) Zinquin fluorescence mossy fiber 目前，检测锌离子的手段很多，其中检测细胞内外游离锌离子的形态学手段主要有两种，即金属自显影术(autometallography, AMG)和；N(6methoxy8quinolyl)ptoluenesulfonamide (TSQ)和Zinquin锌荧光探针技术。Timm创始了著名的硫化银染色法，该方法是将“物理显影”的知识转而应用到重金属