双重标记连接蛋白43激光共聚焦显微镜检测方法的研究

The Method of Detection of Double Labeling Connexin43 by Laser Scanning Confocal Microscope

目的 :探讨激光扫描共聚焦显微镜和荧光双重标记技术对心肌连接蛋白 43进行定量和定位研究的方法。方法 :应用罗丹明 -麦芽凝集素标记心肌细胞膜 ,FITC标记连接蛋白 43;应用激光扫描共聚焦显微镜的检测器 1和检测器 2同时分别检测连接蛋白 43和细胞膜 ,应用Image Analysis程序进行图像分析。结果 :在激光扫描共聚焦显微镜下 ,可以清晰地观察连接蛋白 43和细胞膜。应用Image Analysis程序能对连接蛋白 43进行定量研究 ,连接蛋白 43的分布也能同时得到研究。结论 :结合应用激光扫描共聚焦显微镜和双重标记技术能对连接蛋白 43进行精确的定量和定位研究 。

Objective: To explore a method of detecting the connexin 43(Cx43) quantity and distribution by laser scanning confocal microscope (LSCM) with a double label technique. Methods: The FITC was used to label the Cx43, and the rhodamine(Wheat germ agglutin conjugated) was used to label the sarcolemma. Then, the Cx43 and the sarcolemma were detected separately by the detector 1 and the detector 2 of the SLCM simultaneously, and images were analyzed by the Image Analysis program. Results: The distinct images of the Cx43 and the sarcolemma could be obtained by the LSCM. The Cx43 could be quantitated by the Image-Analysis program, and the distribution of Cx43 could be observed simultaneously. Conclusion: The combination of LSCM and double-label technique is a advanced method of detecting the connexin 43(Cx43) quantity and distribution.