Determining the target protein localization in 3D using the combination of FIB-SEM and APEX2
作者: Yang ShiLi WangJianguo ZhangYujia ZhaiFei Sun
作者单位: 1Chinese Academy of Sciences
2University of Chinese Academy of Sciences
3Sino-Danish Center for Education and Research
刊名: Biophysics Reports, 2017, Vol.3 (4-6), pp.92-99
来源数据库: Springer Nature Journal
DOI: 10.1007/s41048-017-0043-x
关键词: Enhanced ascorbate peroxidaseFocus ion beam scanning electron microscopyMitochondrial dynamicsProtein locationThree-dimensional space
原始语种摘要: Determining the cellular localization of proteins of interest at nanometer resolution is necessary for elucidating their functions. Besides super-resolution fluorescence microscopy, conventional electron microscopy (EM) combined with immunolabeling or clonable EM tags provides a unique approach to correlate protein localization information and cellular ultrastructural information. However, there are still rare cases of such correlation in three-dimensional (3D) spaces. Here, we developed an approach by combining the focus ion beam scanning electron microscopy (FIB-SEM) and a promising clonable EM tag APEX2 (an enhanced ascorbate peroxidase 2) to determine the target protein localization within 3D cellular ultrastructural context. We further utilized this approach to study the 3D...
全文获取路径: Springer Nature  (合作)

  • localization 定位
  • microscopy 显微镜检查
  • protein 蛋白质
  • SEM Sample Evaluation Method
  • nanometer 毫微米
  • electron 电子
  • ascorbate 抗坏血酸
  • promising 前提
  • target 目标
  • scanning 扫描