Sensitivity in horseradish peroxidase neurohistochemistry: a comparative and quantitative study of nine methods.
Sensitivity in horseradish peroxidase neurohistochemistry: a comparative and quantitative study of nine methods.
复制标题
辣根过氧化物酶神经组织化学的敏感性：九种方法的比较和定量研究。
DOI：
10.1177/27.3.113450
复制
发表时间：
1979
期刊：
影响因子：
--
通讯作者：
D. Rosene
中科院分区：
文献类型：
--
作者：
M. Mesulam;D. Rosene
已在四只大鼠和四只恒河猴的匹配组织切片上对九种目前可用的 HRP 神经组织化学方法进行了相互比较。本报告中研究的九种方法是 LaVail JH 和 LaVail MM (J Comp Neurol 157:303, 1974)、Adams JC (Neuroscience 2:141, 1977) 以及 Streit P 和 Reubi JC (Brain Res 126:530, 1977) 的二氨基联苯胺 (DAB) 程序； Mesulam M-M (J Histochem Cytochem 24:1273, 1976)和 De Olmos J 和 Heimer L (Neurosci Lett 6:107, 1977)的二联苯胺二盐酸盐(BDHC)程序； De Olmos J 的邻二茴香胺 (O-D) 程序 (Exp Brain Res 29:541, 1977)； Hanker JS 等人 (Histochem J 9:789, 1977) 的对苯二胺二盐酸盐和邻苯二酚 (PPD-PC) 方法以及 Mesulam M-M (J Histochem Cytochem 26:106, 1978) 和 De Olmos J 等人的四甲基联苯胺 (TMB) 方法(J Comp Neurol 181:213, 1978)。定量比较基于逆行标记的核周体的计数。顺行运输的程度和注射部位的大小也在更定性的水平上进行了比较。结果表明，一种 TMB 程序（Mesulam M-M，J Histochem Cytochem 26：106，1978）在其可以证明的标记核周体数量方面明显优于其他八种程序中的每一种。此外，这些差异在统计上显着，且置信水平优于 0.05。当核周体含有少量转运的 HRP 时，敏感性差异最为明显。与所有其他程序相比，相同的 TMB 方法还展示了更多的顺行运输和更大的注射部位。如果采用不太敏感的程序，则由该 TMB 程序清楚证明的传入或传出连接要么被低估，要么完全被忽视。建议 HRP 神经组织化学的敏感性由多种因素决定，包括固定方法、固定后储存、色原的选择、孵育参数、施用的 HRP 酶类型和反应后处理。
Nine currently available methods for HRP neurohistochemistry have been compared with each other on matching tissue sections from four rats and four rhesus monkeys. The nine methods investigated in this report are the diaminobenzidine (DAB) procedures of LaVail JH and LaVail MM (J Comp Neurol 157:303, 1974), of Adams JC (Neuroscience 2:141, 1977) and of Streit P and Reubi JC (Brain Res 126:530, 1977); the benzidine dihydrochloride (BDHC) procedures of Mesulam M-M (J Histochem Cytochem 24:1273, 1976) and of De Olmos J and Heimer L (Neurosci Lett 6:107, 1977); the o-dianisidine (O-D) procedure of De Olmos J (Exp Brain Res 29:541, 1977); the p-phenylenediamine dihydrochloride and pyrocatechol (PPD-PC) procedure of Hanker JS et al., (Histochem J 9:789, 1977) and the tetramethyl benzidine (TMB) procedures of Mesulam M-M (J Histochem Cytochem 26:106, 1978) and of De Olmos J et al. (J Comp Neurol 181:213, 1978). Quantitative comparisons were based on counts of retrogradely labeled perikarya. The extent of anterograde transport and the size of the injection site were also compared at a more qualitative level. The results indicate that one TMB procedure (Mesulam M-M, J Histochem Cytochem 26:106, 1978) is distinctly superior to each of the other eight procedures in the number of labeled perikarya that it can demonstrate. Furthermore, these differences are statistically significant at better than the 0.05 level of confidence. Differences in sensitivity are most evident when the perikarya contain small quantities of transported HRP. The same TMB method also demonstrates more anterograde transport and a larger injection site than all the other procedures. If less sensitive procedures are employed, afferent or efferent connections that are clearly demonstrated by this TMB procedure are either underestimated or completely overlooked. It is suggested that sensitivity in HRP neurohistochemistry is determined by multiple factors which include the method of fixation, post-fixation storage, the choice of chromogen, the incubation parameters, the type of HRP enzyme that is administered, and the postreaction treatment.