Fluorescence imaging using a fluorescent protein with a large Stokes shift.
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.
Kogure
Takako
T
Laboratory for Cell Function and Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
Kawano
Hiroyuki
H
Abe
Yukiko
Y
Miyawaki
Atsushi
A
D000818:Animals; D002522:Chlorocebus aethiops; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D010957:Plasmids; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism / Q000528:radiation effects; D012107:Research Design; D014162:Transfection; D014709:Vero Cells; D000097573:Red Fluorescent Protein
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C485182:Cyan Fluorescent Protein