使用HyPer2生物传感器进行活细胞受激发射损耗成像

Live-Cell STED Imaging with the HyPer2 Biosensor.

作者信息

Mishina Natalia M, Belousov Vsevolod V

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, Moscow, 117997, Russia.

出版信息

Methods Mol Biol. 2017;1663:21-28. doi: 10.1007/978-1-4939-7265-4_3.

DOI:10.1007/978-1-4939-7265-4_3

PMID:28924656

Abstract

Stimulated emission depletion (STED) microscopy is a popular super resolution imaging technique. Not only synthetic dyes and fluorescent proteins can be utilized as STED fluorophores, but also genetically encoded biosensors. Fusing the biosensor with proteins of interest allows subdiffraction imaging of intracellular macromolecular architecture with simultaneous extraction of functional information about cellular activities. Here, we describe a protocol for live-cell STED microscopy of the HyPer2 biosensor fused to cytoskeletal filaments.

摘要

受激发射损耗（STED）显微镜是一种流行的超分辨率成像技术。不仅合成染料和荧光蛋白可作为STED荧光团，而且基因编码的生物传感器也可以。将生物传感器与感兴趣的蛋白质融合，能够对细胞内大分子结构进行亚衍射成像，同时提取有关细胞活动的功能信息。在此，我们描述了一种用于对与细胞骨架丝融合的HyPer2生物传感器进行活细胞STED显微镜成像的方案。