Maggiano N, Larocca L M, Piantelli M, Ranelletti F O, Lauriola L, Ricci R, Capelli A
Department of Pathology, Università Cattolica del S. Cuore, Rome, Italy.
Histochem J. 1991 Feb;23(2):69-74. doi: 10.1007/BF01047110.
A full-length cDNA clone encoding the constant region of T cell receptor beta chain was labelled by random priming DNA with digoxigenin-dUTP. The probe was used to estimate the relative amount of the receptor beta chain mRNA by in situ hybridization on frozen sections from human thymus and lymph nodes. The hybridization was visualized in blue using an anti-digoxigenin antibody conjugated with alkaline phosphatase and a subsequent enzyme-catalysed colour reaction. The distributions of the signal in tissue sections were as expected. Moreover, labelled cells showed hybrids both in the cytoplasm and in the nucleus, and strongly and weakly stained cells were clearly distinguishable. The results indicate that this method of in situ hybridization should be useful in the detection of specific mRNA in frozen sections.
通过用洋地黄毒苷-dUTP随机引物DNA标记编码T细胞受体β链恒定区的全长cDNA克隆。该探针用于通过对人胸腺和淋巴结冰冻切片进行原位杂交来估计受体β链mRNA的相对量。使用与碱性磷酸酶偶联的抗洋地黄毒苷抗体和随后的酶催化显色反应，以蓝色显示杂交情况。组织切片中信号的分布符合预期。此外，标记细胞在细胞质和细胞核中均显示出杂交体，并且强染色和弱染色的细胞清晰可辨。结果表明，这种原位杂交方法在冰冻切片中特异性mRNA的检测中应是有用的。
