中文摘要：

采用标记线粒体的荧光质粒pDsRed2-Mito转染PC12细胞,分别以95%乙醇、甲醇、丙酮和4%多聚甲醛作为固定剂对筛选后的细胞进行固定,最后在激光共聚焦显微镜下观察不同固定剂对荧光质粒荧光强度的影响.结果表明,95%乙醇或甲醇作为固定剂固定细胞后易导致荧光猝灭,而丙酮或4%多聚甲醛固定细胞后,对质粒荧光强度没有显著影响.不同传代次数对pDsRed2-Mito质粒的荧光强度也没有显著影响.因此,在选择pDsRed2-Mito质粒标记线粒体时,不用考虑传代次数的影响,宜选择丙酮或4%多聚甲醛作为固定剂才能最大化的保持质粒的荧光强度.

英文摘要：

In this paper, the pDsRed2-Mito plasmid, which specifically marks the mitochondrial morphology and structure in cells, was transfected to PC12 cells, and fixed by 95% ethanol, methyl alcohol, acetone and 4% paraformaldehyde respectively. The effect of different fixatives on fluorescence intensity of the plasmid was observed with laser scanning confocal microscope. The results indicate that the fluorescence intensity of pDsRed2-Mito would quench easily when the fixative was 95% ethanol or methyl alcohol, while the introduction of the acetone or 4% paraformaldehyde has no remarkable effect on its fluorescence intensity. Furthermore, the times of passages has also no significant impact on the fluorescence intensity of pDsRed2-Mito. In this way, with pDsRed2-Mito to label mitochondria, it is a good choose to use acetone and 4%paraformaldehyde as a fixative to maintain fluorescence intensity.