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| LCM联合快速免疫组化技术——一种获取肿瘤原位血管内皮细胞的可行途径 | |
| 引用本文： | 陶思,汤多壮,白向阳,孙立石,马丁,孙汉英,周剑峰.LCM联合快速免疫组化技术——一种获取肿瘤原位血管内皮细胞的可行途径[J].中国组织化学与细胞化学杂志,2009,18(1). |
| 作者姓名： | 陶思 汤多壮 白向阳 孙立石 马丁 孙汉英 周剑峰 |
| 作者单位： | 1. 华中科技大学同济医学院附属同济医院血液科,武汉,430030 2. 首都医科大学附属北京佑安医院肝病 3. 华中科技大学同济医学院附属同济医院,肿瘤生物医学中心,武汉,430030 4. 华中科技大学同济医学院附属同济医院血液科,武汉,430030;华中科技大学同济医学院附属同济医院,肿瘤生物医学中心,武汉,430030 |
| 摘 要： | 目的肿瘤血管内皮细胞对肿瘤发生发展极为重要,是目前肿瘤研究的热点。本研究为从肿瘤组织原位获取高纯度血管内皮细胞进行基因表达研究摸索可行方法。方法获取淋巴瘤组织标本后,置于锌固定液中固定,并通过进行激光捕获显微切割(lasercapture microdissection,LCM)前操作模拟LCM环境,确定锌固定法对RNA完整性的保护作用。将组织标本制作冰冻切片,采用快速免疫组化染色方法标记血管内皮细胞,利用LCM技术获取肿瘤组织原位血管内皮细胞,并用RT-PCR方法对所获细胞进行纯度检测。结果无论是固定后直接提取组织RNA,还是经模拟LCM环境再提取RNA,均显示锌固定法对RNA完整性提供了良好保护。快速免疫组化可以明确标记血管内皮细胞,后者能够被LCM准确捕获,并经RT-PCR验证为高纯度的血管内皮细胞。结论快速免疫组化联合LCM技术可以从肿瘤组织原位获取高纯度血管内皮细胞,并保证RNA的完整性,可能为肿瘤血管内皮细胞基因表达研究奠定基础。 |
| 关 键 词： | 肿瘤 血管内皮细胞 激光捕获显微切割 快速免疫组化 基因芯片 |
COMBINATION OF QUICK STAINING IMMUNOHISTOCHEMISTRY AND LASER CAPTURE MICRO-DISSECTION-A FEASIBLE APPROACH TO OBTAINING HIGHLY PURE TUMOR VASCULAR ENDOTHLIAL CELLS FROM IN SITU TUMOR TISSUES |
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| Tao Si,Tang Duozhuang,Bai Xiangyang,Sun Lishi,Ma Ding,Sun Hanying,Zhou Jianfeng.COMBINATION OF QUICK STAINING IMMUNOHISTOCHEMISTRY AND LASER CAPTURE MICRO-DISSECTION-A FEASIBLE APPROACH TO OBTAINING HIGHLY PURE TUMOR VASCULAR ENDOTHLIAL CELLS FROM IN SITU TUMOR TISSUES[J].Chinese Journal of Histochemistry and Cytochemistry,2009,18(1). | |
| Authors: | Tao Si Tang Duozhuang Bai Xiangyang Sun Lishi Ma Ding Sun Hanying Zhou Jianfeng |
| Institution: | 1Department of Hematology;Tongji Hospital;Tongji Medical College;Huazhong University of Science and Technology;Wuhan 430030;China;2The 5th Department of Hepatopathy;Beijing Youan Hospital;Capital Medical University;Beijing 100069;3Cancer Biology Research Center;China |
| Abstract: | Objective To develope a feasible approach of obtaining tumor vascular endothelial cells of high purity from in situ tumor tissues.Methods Clinical specimens were fixed in the zinc fixation fluid before RNA extraction, and the integrity of RNA was checked by gel electrophoresis to determine the protective effect of the fluid. Laser capture micro-dissection (LCM) condition was also simulated to examine the protective effect of the fluid for the LCM process. The in situ tumor vascular endothelial cells were pr... |
| Keywords: | Tumor Vascular endothelial cell Laser capture microdissection（LCM） Quick staining immunohistochemistry Gene expression microarray |
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