Liu Yingchao, Wu Jinsong, Liu Sixiu, Zhuang Dongxiao, Wang Yongfei, Shou Xuefei, Zhu Jianhong
Shanghai Neurosurgical Center, Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
Colloids Surf B Biointerfaces. 2009 Jul 1;71(2):187-93. doi: 10.1016/j.colsurfb.2009.02.005. Epub 2009 Feb 20.
Laser capture microdissection (LCM) technology combined with immunohistochemistry (immuno-LCM) is a valuable tool to obtain specific target cell populations and therefore this technique enables more accurate proteomic profile. In this study, we optimized the regular immuno-LCM technique to isolate and stain pure prolactin cells from either normal human pituitary (n=6) or prolactioma (n=11). Compared with the routine procedure, more intense and specific staining could be obtained when sections were pretreated with 0.2% Triton X-100 for 4 min. Interestingly, longer pretreatment (0.2% Triton X-100 for 10 min) or higher concentration (2% Triton X-100 for 4 and 10 min) greatly impaired labeling intensity and cell shape. Further scanning electron microscope study revealed that the component extracted from the cell surface by Triton X-100 was lipid. Using the optimized immuno-LCM technique, more pure prolactin cells could be isolated and prepared for further proteomic analysis. Taken together, we reported an optimized immuno-LCM technique that could effectively dissect pure target cells in different type pituitary adenomas for further proteomics analysis.
激光捕获显微切割（LCM）技术与免疫组织化学相结合（免疫LCM）是获取特定靶细胞群体的一种有价值的工具，因此该技术能够实现更准确的蛋白质组学分析。在本研究中，我们优化了常规免疫LCM技术，以从正常人垂体（n = 6）或催乳素瘤（n = 11）中分离并染色纯催乳素细胞。与常规程序相比，当切片用0.2% Triton X-100预处理4分钟时，可以获得更强且更特异的染色。有趣的是，更长时间的预处理（0.2% Triton X-100处理10分钟）或更高浓度（2% Triton X-100处理4分钟和10分钟）会极大地损害标记强度和细胞形态。进一步的扫描电子显微镜研究表明，Triton X-100从细胞表面提取的成分是脂质。使用优化的免疫LCM技术，可以分离出更多纯催乳素细胞，并为进一步的蛋白质组学分析做好准备。综上所述，我们报道了一种优化的免疫LCM技术，该技术可以有效地在不同类型垂体腺瘤中分离出纯靶细胞，用于进一步的蛋白质组学分析。
