Würden S, Homberg U
Fakultät für Biologie der Universität, Konstanz 1, Germany.
J Histochem Cytochem. 1993 Apr;41(4):627-30. doi: 10.1177/41.4.8450202.
We have developed a new double immunofluorescence technique by which two neuroactive substances in the same tissue section can be labeled with primary antisera raised in the same species. The optic lobes of the locust Schistocerca gregaria were used as a model system to develop the staining procedure. FMRFamide-immunoreactive neurons were detected by rabbit antisera against FMRFamide and FITC-conjugated secondary antibodies. Antibodies against the second peptide, pigment-dispersing hormone (PDH), also raised in rabbit, were biotinylated and detected via streptavidin-Texas Red. Crossreactivity of the PDH immunoglobulins with the FITC-conjugated secondary antiserum was prevented by pre-incubation with rabbit gamma globulins. The two peptide immunoreactivities could be conveniently observed on the same section with the different fluorescent markers. This double labeling technique with modified antibodies is easily performed and highly useful for co-localization studies with antisera raised in the same species.
我们开发了一种新的双重免疫荧光技术，通过该技术可以用在同一物种中产生的一抗标记同一组织切片中的两种神经活性物质。以沙漠蝗（Schistocerca gregaria）的视叶作为模型系统来开发染色程序。用抗FMRF酰胺的兔抗血清和异硫氰酸荧光素（FITC）偶联的二抗检测FMRF酰胺免疫反应性神经元。同样在兔中产生的针对第二种肽——色素分散激素（PDH）的抗体用生物素进行标记，并通过链霉亲和素-德克萨斯红进行检测。通过与兔γ球蛋白预孵育来防止PDH免疫球蛋白与FITC偶联的二抗之间的交叉反应。可以用不同的荧光标记在同一切片上方便地观察到两种肽的免疫反应性。这种用修饰抗体的双重标记技术易于操作，对于使用在同一物种中产生的抗血清进行共定位研究非常有用。
