Fluorescence imaging using a fluorescent protein with a large Stokes shift

Methods. 2008 Jul;45(3):223-6. doi: 10.1016/j.ymeth.2008.06.009. Epub 2008 Jun 27.

Abstract

Keima is a far-red fluorescent protein endowed with a large Stokes shift. It absorbs light maximally at around 440nm and emits maximally at around 620nm. While the original Keima is obligately tetrameric (tKeima), the dimeric and monomeric versions (mKeima and dKeima, respectively) have been generated. More recently, a tandem dimer of Keima (tdKeima) has been developed as the brightest version. Here we describe examples, which show the usefulness of Keima for dual-color fluorescence imaging technologies, such as fluorescence cross-correlation spectroscopy (FCCS) and two-photon laser scanning microscopy (TPLSM). Keima can be used in conjunction with existing fluorescent proteins in which the Stokes shift is much smaller, with the idea that while two fluorescent proteins are excited by a single laser each will fluoresce a different color.

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Luminescent Proteins / analysis*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Plasmids
  • Protein Engineering
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / radiation effects
  • Red Fluorescent Protein
  • Research Design
  • Transfection
  • Vero Cells

Substances

  • Cyan Fluorescent Protein
  • Fluorescent Dyes
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins