激光扫描共聚焦显微镜中荧光强度测定条件探究.docVIP

激光扫描共聚焦显微镜中荧光强度测定条件探究 　　　【摘要】 目的 本文对应用激光扫描共聚焦显微镜进行荧光强度的测定条件做了详细的分析。方法 选择和设置激光扫描共聚焦显微镜的各参数，对用间接免疫荧光法检测改性后的材料表面对人牙龈成纤维细胞分泌细胞外基质纤粘连蛋白FN和I型胶原的影响进行荧光强度的定量。结果 细胞在材料表面接种后FN的荧光染色强度在四组材料之间差异无显著性，说明材料表面化学成分的改变对FN的形成没有明显影响。但四组材料对I型胶原分泌的影响有差异。结论 选择和设置适合的激光扫描共聚焦显微镜中荧光强度的测定条件，可以真实有效的反映实验的结果。 　　关键词 激光扫描共聚焦显微镜 荧光强度 　　 　　The research on determine condition of fluorescence intensity in confocal laser scanning microscopy 　 　　【Abstract】 Objective To explore the conditions under which fluorescence intensity is determined in confocal laser scanning microscopy.Methods The parameters of confocal laser scanning microscopy were set and the effects of these modifications on the response of the fibro-conglutination albumen and type I collage n of human gingival fibrobˉlasts on indirect immunityfluorescence were observed，and the fluorescence intensity of the effects was measured.Reˉsults After the cells grafted on the material surface，the fluorescence dye intensity of the fibro-conglutination albuˉmen had no notable differ ence in materials of the four groups，the alter of the chemistry component of the material surˉface had no notable influence on the formation of the fibro-conglutination albumen.But the effects of the type I collaˉgen were different in materials of the four groups.Conclusion Setting appropriate conditions for the determination of fluorescence intensity in confocal laser scanning microscopy is important in the analysis of the true results of experiˉment.Key words confocal laser scanning microscopy fluorescence intensity 　　激光扫描共聚焦显微镜（confocal laser scanning miˉcroscopy，CLSM）是20世纪80年代发展起来的目前世界上最先进的分子细胞生物学分析仪器。它是在荧光显微镜基础上加装了激光扫描装置，使用紫外或可见激光激发荧光探针，从而得到细胞或组织内部微细结构的荧光图像，较传统显微镜有着不可比拟的优越性 ［1］ ，已广泛应用于分子细胞生物学、生理学、病理学、免疫学、药理学、遗传学等领域，成为这些领域新一代强有力的研究工具，对生物样品进行定性、定量、定时和定位研究具有很大的优越性 。 　　1 材料与方法 　　1.1 实验用样品 分四组。Ti组:未涂层的对照组;HA组:应用离子束辅助沉积技术在种植体表面沉积羟基磷灰石HA涂层;TCP/HA组:应用离子束辅助沉积技术在种植体表面沉积磷酸三钙/羟基磷灰石TCP/HA复合涂层;RGD组:应用化学方法在纯钛表面共价接枝生物活性肽RGD（Arg-Gly-Asp，精氨酸-甘氨酸-天门冬氨酸）。样品规格:直径10mm，厚2mm的圆盘形，表