激光捕获显微切割制备垂体腺瘤蛋白质组学研究样品

目的 探讨应用免疫组织化学导航激光捕获显微切割(LCM)技术获取高同质性泌乳素垂体腺瘤细胞及正常垂体泌乳素细胞的方法及可行性.方法 冰冻泌乳素腺瘤标本及对照正常垂体常规制备8μm冰冻覆膜切片,采用两步法免疫组织化学方法染色;应用LCM技术切割腺瘤内及正常对照垂体内泌乳素细胞;选择性获取同质泌乳素细胞和配对正常细胞.结果 60 min内完成免疫组织化学染色,细胞显色较理想.采用4μm紫外线激光可以准确、快速地分离片状甚至单个泌乳素分泌细胞;无间质细胞污染,细胞形态保持完好.结论 应用LCM技术可以成功获垂体瘤蛋白组学研究中的异质性问题,所获组织同质性高、操作过程稳定是垂体瘤蛋白质组学研究较有价值的样品制备方法.

Application of immunohistochemistry navigated laser capture microdissection to capture pure prolactin cells for proteomics

Objective To investigate the effect of immunohistochemistry navigated laser capture microdissection (LCM) to prepare pure targeted cells from prolactin pituitary adenoma and normal pituita-ry for proteomic research. Methods Immunohistochemistry navigated-LCM technique was employed to successfully obtain the pure cluster or even single cell from prolactnoma and normal pituitary frozen sec-tions. Results Immunohistochemistry could be achieved within 60 min, and 4 μm ultraviolet laser was ap-plied to quickly and precisely obtain pure targeted cells with optimal morphology but without interstitium pollution. Conclusion Immunohistochemistry navigated LCM was a valuable technique which can tackle the problem of tissue heterogeneity in proteomic research with high homogeneity and stable operative proce-dure.