Beckers Patrick, Müller Claudia, Wallnisch Christiane, Bartolomaeus Thomas
Institute of Evolutionary Biology, University of Bonn, An der Immenburg 1 53121 Bonn, Germany.
J Biol Methods. 2021 Sep 3;8(3):e153. doi: 10.14440/jbm.2021.354. eCollection 2021.
Classical histological stained sections have the disadvantage that fine structures, like individual neurites, or specific macromolecules, like neurotransmitters cannot be visualized. Due to its highly specific staining of only one target molecule within the cell, the visualization of delicate structures, which would be superimposed by other tissue layers in classical Azan staining, is possible with immunohistochemistry. However, using immunohistological methods not all tissues of a specimen can be visualized at once. In contrast, density specific stains like Azan allow for a whole staining of the tissues. We provide a step by step protocol of how to combine immunohistochemistry and Azan staining in the same serial paraffin sections. The combination of both methods allows for a highly detailed investigation of structures of interest. The spatial detection of the previous, to Azan staining, gained antibody-labeled signal allows for a much better understanding of animal organ systems. By using serial sections, it is possible to create an aligned image stack that is both Azan stained and also antibody-labeled. Thus enabling a correlative approach that bridges traditional histology with immunohistochemistry in animal morphology.
传统组织学染色切片的缺点是，像单个神经突这样的精细结构或像神经递质这样的特定大分子无法可视化。由于免疫组织化学仅对细胞内的一种靶分子进行高度特异性染色，因此可以可视化在传统偶氮染色中会被其他组织层叠加的精细结构。然而，使用免疫组织学方法不能同时可视化标本的所有组织。相比之下，像偶氮染色这样的密度特异性染色可以对组织进行整体染色。我们提供了一个在同一连续石蜡切片中如何结合免疫组织化学和偶氮染色的分步方案。两种方法的结合可以对感兴趣的结构进行高度详细的研究。对偶氮染色之前获得的抗体标记信号进行空间检测，可以更好地理解动物器官系统。通过使用连续切片，可以创建一个既进行了偶氮染色又进行了抗体标记的对齐图像堆栈。从而实现一种在动物形态学中将传统组织学与免疫组织化学联系起来的相关方法。
