Coghlan J P, Aldred P, Haralambidis J, Niall H D, Penschow J D, Tregear G W
Anal Biochem. 1985 Aug 15;149(1):1-28. doi: 10.1016/0003-2697(85)90472-5.
In this review we have used our own recent work as a flagship to illustrate the recent renaissance of interest in hybridization histochemistry. A trickle of papers followed the initial key excursion into the in situ labeling of tissue sections (48-50). Our own entry into this field started in 1978 and since then a confluence of important questions and technical advances has served to make hybridization histochemistry much more attractive as a research tool. Hybridization histochemistry is able to solve some problems for which there is no other suitable technique at this time. Hybridization histochemistry provides the location of anatomical sites of gene expression, and viral replication, with uniquely high specificity. We have taken 32P-labeled probes to what appears to be their limit of resolution, which is single cells in thin sections. While 32P has clear disadvantages, exposure time is relatively short and the use of fast-X-ray film to preview the results and estimate exposure time for emulsion has been turned to advantage. Our introduction (27) of the use of whole-mouse sections in hybridization histochemistry has great potential in hormonal, enzymatic, and growth factor gene expression and will no doubt prove of great use in developmental studies and examination of viral infection. The use of synthetic DNA (synthetic oligonucleotides) unshackles the technique from the need for an associated molecular biology laboratory and at once widens the horizon of application of the technique. Although hybridization histochemistry is a valuable research tool which will soon find a niche in many fields, in a short time it should become a key diagnostic aid. It may well become the method of preference for detection of the expression of oncogenes and other cancer-related genes and for viruses which for other reasons are difficult to detect.
在本综述中,我们以自己最近的工作为范例,来说明对杂交组织化学兴趣的再度兴起。在最初对组织切片原位标记进行关键探索之后,陆续有少量论文发表(48 - 50)。我们自己进入这个领域始于1978年,从那时起,一系列重要问题和技术进步使得杂交组织化学作为一种研究工具更具吸引力。杂交组织化学能够解决一些目前尚无其他合适技术可解决的问题。杂交组织化学能以极高的特异性确定基因表达和病毒复制的解剖学位置。我们已将32P标记探针的分辨率发挥到似乎是其极限,即在薄切片中达到单细胞水平。虽然32P有明显缺点,但曝光时间相对较短,利用快速X射线胶片预览结果并估计对乳胶的曝光时间已发挥了优势。我们在杂交组织化学中引入全鼠切片的方法(27)在激素、酶和生长因子基因表达方面有很大潜力,无疑将在发育研究和病毒感染检测中证明有很大用途。合成DNA(合成寡核苷酸)的使用使该技术摆脱了对相关分子生物学实验室的需求,立即拓宽了该技术的应用范围。尽管杂交组织化学是一种有价值的研究工具,很快会在许多领域找到用武之地,但在短时间内它应该会成为一种关键诊断辅助手段。它很可能会成为检测癌基因和其他癌症相关基因表达以及检测因其他原因难以检测的病毒的首选方法。