Hoffmann F, Janussen D, Dröse W, Arp G, Reitner J
University of Göttingen, Department of Geosciences, Geobiology Group, Goldschmidtstrasse 3, D-37077 Göttingen, Germany.
Biotech Histochem. 2003 Jun-Aug;78(3-4):191-9. doi: 10.1080/10520290310001613042.
Siliceous and calcareous sponges commonly are treated with acid to remove the spicules prior to embedding and cutting for histological investigations. Histology of spiculated sponge tissue represents a challenging problem in sponge histotechnology. Furthermore, fluorescence in situ hybridization (FISH), a key method for studying sponge-associated microbes, is not possible after acid treatment. For a broad range of siliceous sponge species, we developed and evaluated methods for embedding in paraffin, methylmethacrylate resins, LR White resin and cryomatrix. Different methods for cutting tissue blocks as well as mounting and staining sections also were tested. Our aim was to enable histological investigations and FISH without prior removal of the spicules. To obtain an overview of tissue and skeleton arrangement, we recommend embedding tissue blocks with LR White resin combined with en bloc staining techniques for large specimens with thick and numerous spicules, but paraffin embedding and subsequent staining for whole small specimens. For FISH on siliceous sponges, we recommend Histocryl embedding if the spicule content is high, but paraffin embedding if it is low. Classical histological techniques are used for detailed tissue examinations.
在进行组织学研究而进行包埋和切片之前，硅质海绵和钙质海绵通常要用酸处理以去除骨针。有骨针的海绵组织的组织学在海绵组织技术中是一个具有挑战性的问题。此外，酸处理后无法进行荧光原位杂交（FISH），这是研究与海绵相关微生物的关键方法。对于广泛的硅质海绵物种，我们开发并评估了用石蜡、甲基丙烯酸甲酯树脂、LR White树脂和冷冻基质进行包埋的方法。还测试了切割组织块以及装片和染色切片的不同方法。我们的目标是在不事先去除骨针的情况下进行组织学研究和FISH。为了了解组织和骨骼的排列情况，对于有粗大且数量众多骨针的大型标本，我们建议用LR White树脂结合整体染色技术包埋组织块，但对于整个小型标本则采用石蜡包埋和后续染色。对于硅质海绵的FISH，如果骨针含量高，我们建议用Histocryl包埋，如果含量低，则建议用石蜡包埋。经典的组织学技术用于详细的组织检查。
