Rogers Sally, Scholpp Steffen
Living Systems Institute, School of Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, UK.
Methods Mol Biol. 2021;2346:183-190. doi: 10.1007/7651_2020_305.
Cytonemes are specialized signalling filopodia that have a role in development and cellular differentiation. However, they are not well preserved by standard fixation techniques to study protein localization and interactions. A recent methodological advance has yielded improvements in cytoneme preservation using glutaraldehyde fixation and sodium borohydride treatment to reduce background. We herein describe a safer method for effective blocking using glycine following glutaraldehyde fixation of cytonemes on cultured adherent cells and demonstrate its effectiveness in immunocytochemistry.
细胞触须是一种特殊的信号丝状伪足，在发育和细胞分化中发挥作用。然而，标准固定技术并不能很好地保存它们，以便研究蛋白质定位和相互作用。最近的一项方法学进展通过使用戊二醛固定和硼氢化钠处理来减少背景，从而在细胞触须保存方面取得了改进。我们在此描述一种更安全的方法，即在对培养的贴壁细胞上的细胞触须进行戊二醛固定后，使用甘氨酸进行有效封闭，并证明其在免疫细胞化学中的有效性。
