Centre for Organismal Studies, University of Heidelberg, Heidelberg, Baden-Württemberg, Germany.
PLoS One. 2011;6(5):e19713. doi: 10.1371/journal.pone.0019713. Epub 2011 May 13.
For the detection and sub-cellular (co)-localization of proteins in the context of the tissue or organism immunostaining in whole mount preparations or on sections is still the best approach. So far, each antibody required its own fixation and antigen retrieval protocol so that optimizing immunostaining turned out to be tedious and time consuming.
METHODOLOGY/PRINCIPAL FINDING: Here we present a novel method to efficiently retrieve the antigen in a widely applicable standard protocol, facilitating fluorescent immunostaining of both cryosections and whole mount preparations in zebrafish (Danio rerio) and medaka (Oryzias latipes).
CONCLUSIONS/SIGNIFICANCE: Our method overcomes the loss of sections and damage of tissue and cell morphology, and allows parallel immunostaining in multiple colors, co-immunostaining with fluorescent proteins in transgenic fish lines and in combination with whole mount in situ hybridization.
在组织或生物体的背景下检测和亚细胞（共）定位蛋白质，免疫染色仍然是最好的方法。到目前为止，每种抗体都需要自己的固定和抗原检索方案，因此优化免疫染色结果变得繁琐且耗时。
方法/主要发现：本文提出了一种新的方法，可以在广泛适用的标准方案中有效地检索抗原，促进斑马鱼（Danio rerio）和青鳉（Oryzias latipes）的冰冻切片和全组织免疫荧光染色。
结论/意义：我们的方法克服了切片丢失和组织损伤及细胞形态破坏的问题，并且允许在多个颜色中进行平行免疫染色，与转基因鱼系中的荧光蛋白共免疫染色，并与全组织原位杂交相结合。
