https://scholars.lib.ntu.edu.tw/handle/123456789/606886
標題: | Surface charge manipulation and electrostatic immobilization of synaptosomes for super-resolution imaging: a study on tau compartmentalization | 作者: | Bhattacharya U Jhou J.-F Zou Y.-F Abrigo G Lin S.-W Chen Y.-H Chien F.-C Tai H.-C. HWAN-CHING TAI |
關鍵字: | synaptophysin;tau protein;animal;fluorescence microscopy;metabolism;mouse;postsynaptic density;procedures;static electricity;synapse;synaptosome;Animals;Mice;Microscopy, Fluorescence;Post-Synaptic Density;Static Electricity;Synapses;Synaptophysin;Synaptosomes;tau Proteins | 公開日期: | 2021 | 卷: | 11 | 期: | 1 | 來源出版物: | Scientific Reports | 摘要: | Synaptosomes are subcellular fractions prepared from brain tissues that are enriched in synaptic terminals, widely used for the study of neural transmission and synaptic dysfunction. Immunofluorescence imaging is increasingly applied to synaptosomes to investigate protein localization. However, conventional methods for imaging synaptosomes over glass coverslips suffer from formaldehyde-induced aggregation. Here, we developed a facile strategy to capture and image synaptosomes without aggregation artefacts. First, ethylene glycol bis(succinimidyl succinate) (EGS) is chosen as the chemical fixative to replace formaldehyde. EGS/glycine treatment makes the zeta potential of synaptosomes more negative. Second, we modified glass coverslips with 3-aminopropyltriethoxysilane (APTES) to impart positive charges. EGS-fixed synaptosomes spontaneously attach to modified glasses via electrostatic attraction while maintaining good dispersion. Individual synaptic terminals are imaged by conventional fluorescence microscopy or by super-resolution techniques such as direct stochastic optical reconstruction microscopy (dSTORM). We examined tau protein by two-color and three-color dSTORM to understand its spatial distribution within mouse cortical synapses, observing tau colocalization with synaptic vesicles as well postsynaptic densities. ? 2021, The Author(s). |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85115354054&doi=10.1038%2fs41598-021-98142-1&partnerID=40&md5=904f237f440538cb74567d9ad793fe75 https://scholars.lib.ntu.edu.tw/handle/123456789/606886 |
ISSN: | 20452322 | DOI: | 10.1038/s41598-021-98142-1 |
顯示於: | 化學系 |
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