Jackson P, Dockey D A, Lewis F A, Wells M
Department of Pathology, University of Leeds.
J Clin Pathol. 1990 Oct;43(10):810-2. doi: 10.1136/jcp.43.10.810.
An in situ hybridisation technique that uses 1 nm immunogold reagents and silver enhancement was devised to detect biotinylated DNA viral probes in formalin fixed, paraffin wax sections of human cervix. DNA probes labelled with biotin-11-deoxyuridine triphosphate were detected after hybridisation to nucleic acid sequences by an antibiotin antibody, followed by a gold labelled secondary antibody. Silver enhancement then permitted visualisation of the signal at the light microscopic level. The method was reliable and produced less background staining than previously described methods. The signal could be enhanced by epi polarisation microscopy. Furthermore, biotinylated DNA probes may be detected directly by a 1 nm gold labelled goat antibiotin antibody without loss of labelling intensity, and this may be preferable to the longer two layer technique, previously described.
设计了一种原位杂交技术，该技术使用1纳米免疫金试剂和银增强法来检测人宫颈福尔马林固定石蜡切片中的生物素化DNA病毒探针。用生物素-11-脱氧尿苷三磷酸标记的DNA探针在与核酸序列杂交后，通过抗生物素抗体进行检测，随后是金标记的二抗。然后通过银增强在光学显微镜水平上观察信号。该方法可靠，背景染色比先前描述的方法少。通过落射偏光显微镜可以增强信号。此外，生物素化的DNA探针可以直接用1纳米金标记的山羊抗生物素抗体检测，而不会损失标记强度，这可能比先前描述的较长的两层技术更可取。
