The use of miniSOG in the localization of mitochondrial proteins

Methods Enzymol. 2014:547:165-79. doi: 10.1016/B978-0-12-801415-8.00010-2.

Abstract

In this chapter, we provide details along with considerations and future directions for the use of miniSOG (for mini Singlet Oxygen Generator), a versatile label for correlated light and electron microscopy of genetically tagged proteins in cells, tissues, and organisms. This new visualizable genetic tag improves the ability of biologists to locate specific proteins at nanoscale resolution and to see these tagged proteins in the environment of structural landmarks that we are used to navigating by, such as mitochondrial membranes and compartments. miniSOG provides high-quality ultrastructural preservation and permits three-dimensional protein localization via electron tomography or serial section block-face scanning electron microscopy. miniSOG is now doing for electron microscopy what the family of green fluorescent protein did for fluorescence microscopy.

Keywords: Correlated light and electron microscopy; Genetic tag; Mitochondrial calcium uniporter; Photooxidation; Protein label; miniSOG.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Calcium Channels / analysis
  • Calcium Channels / genetics
  • Calcium Channels / metabolism
  • Cells, Cultured
  • Electron Microscope Tomography / methods*
  • Light
  • Microscopy, Electron / methods*
  • Microscopy, Electron, Scanning / methods
  • Microtomy / methods
  • Mitochondrial Proteins / analysis*
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism*
  • Oxidation-Reduction
  • Phototropins / genetics
  • Phototropins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Rodentia
  • Singlet Oxygen / metabolism*
  • Transfection / methods

Substances

  • Calcium Channels
  • Mitochondrial Proteins
  • Phototropins
  • Recombinant Proteins
  • mitochondrial calcium uniporter
  • Singlet Oxygen