Brubacher John L, Vieira Ana P, Azimzadeh Juliette
Canadian Mennonite University, Winnipeg, MB, Canada.
Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Methods Mol Biol. 2018;1774:519-538. doi: 10.1007/978-1-4939-7802-1_23.
The flatworm Schmidtea mediterranea and related species are emerging model systems in such fields as stem-cell biology, regeneration, and evolutionary biology. Excellent molecular tools have been developed for S. mediterranea, but ultrastructural techniques have received less attention, which is unfortunate, as these methods are necessary to better understand the actual histological, cellular and subcellular features of regeneration and development. Tissue-processing regimens can be quite idiosyncratic for particular species or specimen types-what works for mammalian tissues or cell cultures will not necessarily give good results with freshwater planarians. Here we present detailed, optimized protocols for preparation of S. mediterranea by "standard" chemical fixation, and by a rapid microwave-based technique.
扁形虫地中海涡虫及相关物种正在成为干细胞生物学、再生和进化生物学等领域新兴的模式系统。针对地中海涡虫已开发出了出色的分子工具，但超微结构技术受到的关注较少，这很遗憾，因为这些方法对于更好地理解再生和发育的实际组织学、细胞及亚细胞特征是必不可少的。组织处理方案对于特定物种或标本类型可能非常独特——适用于哺乳动物组织或细胞培养的方法不一定能在淡水涡虫上取得良好效果。在此，我们展示了通过“标准”化学固定以及基于快速微波技术制备地中海涡虫的详细优化方案。
