摘要
目的 :探讨和建立石蜡包埋组织和新鲜冷冻标本制备组织芯片的方法 ,以及在基因表达分析中的应用。方法 :选取肺癌石蜡包埋组织 90例 ,新鲜液氮冻存肺癌及癌旁肺组织 2 0例 ,常规切片HE染色下确定穿刺部位 ,石蜡包埋组织芯片制作按Kononem等方法 (1998) ,每例穿取 0 6mm组织柱 2条 ,制备 180点阵组织芯片 ;而新鲜组织芯片参考Fejzo等方法 (2 0 0 1) ,并略作改良 ,制备 5 0点阵组织芯片。所有组织芯片模块 ,经 4 μm切片、常规组织学染色和免疫组化染色 ,以评价其在形态学及基因表达分析中应用的可行性。结果 :180点阵石蜡包埋肺癌组织芯片 ,组织排列整齐 ,HE染色后组织形态可观察率在 98%以上 ;p5 3、c myc、bcl 2、bax和Ki 6 7等免疫组化染色后 ,肺癌组织可见不同程度的抗原表达 ,组织点阵的形态可观测率在 89%~ 95 %之间。新鲜组织芯片经HE染色后 ,形态可观测率和基因表达可分析率在 70 %左右。结论 :手工法组织芯片制备简便易行 ,使用组织少 ,不破坏原蜡块 ;实验均一性、可比性好 ,且节省试剂。与常规组织切片相比 ,2点 0 6mm组织可以获得原蜡块 95 %以上的信息 ,完全可以用于组织形态学和蛋白水平上的基因表达分析 ;新鲜组织芯片技术已经初步建立 ,但尚有待于进一步完善制备技术和mRNA分析方法。
Puropose To establish the tissue microarray (TMA) techniques of frozen and paraffin embedded tissues of lung carcinoma and to explore their application in gene expression analyses. Methods A total of 180 0.6 mm samples from 90 paraffin embedded blocks and 50 dot TMA from 25 frozen tissues of lung cancer were constructed in this study. Routine 4 μm sections were cut from the TMA blocks. Haematoxylin eosin and immunohistochemical stains were performed on the sections. Results The dots on TMAs were regularly arranged and their morphology were well preserved. Hamatoxylin eosin staining showed that 98% samples in paraffin section of TMA could be analyzed morphologically. Expression of p53, c myc,bcl 2, bax and Ki 67 were detected in the most dots of TMA sections. Well preserved morphology and immunohistochemical activities were also observed in about 70% dots of the frozen sections. Conclusions Manual method for the preparation of tissue microarray from both fresh and paraffin embedded specimens is an easy and effective technique for analysis of genetic expression in lung cancer. It has the advantages of experimental uniformity and decreased assay volume, and does not destroy original block for diagnosis. Analysis of only two histo spots results in >95% accuracy, compared with that using standard tissue sections. Frozen tissue array technique is successfully set up and mRNA analysis of it will be further investigated.
出处
《临床与实验病理学杂志》
CAS
CSCD
2002年第4期404-407,共4页
Chinese Journal of Clinical and Experimental Pathology
基金
全军十五科研基金资助课题 (No 0 1MA0 71)
关键词
肺癌
石蜡包埋组织
新鲜标本组织芯片
制备
方法学
tissue microarray
lung neoplasms
paraffin embedded tissue
fresh tissue
genetic expression