Fortunato Franco, Hackert Thilo, Büchler Markus W, Kroemer Guido
Department of General, Visceral and Transplantation Surgery, University Clinic Heidelberg, Heidelberg, Germany; Section Surgical Research, University Clinic Heidelberg, Heidelberg, Germany.
Department of General, Visceral and Transplantation Surgery, University Clinic Heidelberg , Heidelberg, Germany.
Mol Cell Oncol. 2016 Oct 27;3(6):e1251382. doi: 10.1080/23723556.2016.1251382. eCollection 2016.
For many years it has been believed that ultrastructural analysis by transmission electron microscopy (TEM) is not possible using frozen tissues. We have developed a TEM method that allows the evaluation of organelles using pancreatic tissue that was previously liquid nitrogen snap-frozen and stored long-term at -80°C. This method is suitable for the quantitative assessment of mitochondria, rough endoplasmic reticulum (RER), and Golgi structures, as well as organelles originating from autophagy signaling. Frozen pancreatic tissue exhibited no signs of freezing- or storage-related damage and was undistinguishable from fresh material subjected to standard glutaraldehyde fixation. Since pancreatic tissue is the most delicate tissue to work with due to the high expression of digestive enzymes, our method is also suitable for other tissue types such as liver. Thus, by applying proper tissue freezing and fixation techniques, retrospective TEM analysis can be performed on mammalian tissues in a time- and cost-saving manner.
多年来，人们一直认为使用冷冻组织无法通过透射电子显微镜（TEM）进行超微结构分析。我们开发了一种TEM方法，该方法可使用先前经液氮速冻并在-80°C长期保存的胰腺组织来评估细胞器。该方法适用于对线粒体、粗面内质网（RER）、高尔基体结构以及源自自噬信号的细胞器进行定量评估。冷冻的胰腺组织未表现出与冷冻或储存相关的损伤迹象，与经过标准戊二醛固定的新鲜材料没有区别。由于胰腺组织因消化酶的高表达而成为最难处理的组织，我们的方法也适用于其他组织类型，如肝脏。因此，通过应用适当的组织冷冻和固定技术，可以以节省时间和成本的方式对哺乳动物组织进行回顾性TEM分析。
