Intravital imaging of green fluorescent protein using two-photon laser-scanning microscopy.
Imaging a fluorophore in a living tissue presents several unique problems. The fluorescence from the labeled cell(s) may be weak, the labeled cells may be buried deep within tissue and the presence of a fluorophore may render the cells photo-sensitive. Two-photon laser-scanning microscopy (TPLSM) offers several advantages in meeting these challenges. We show that TPLSM provides greater sensitivity, better resolution and less photo-bleaching, as compared to confocal laser-scanning microscopy. The dramatically reduced photo-bleaching makes it possible to image cells continuously for long periods of time. Therefore, TPLSM allows a safer and higher-resolution means of imaging living cells labeled with a variety of fluorophores, including green fluorescent protein.
Potter
S M
SM
Beckman Institute (139-74), Division of Biology, California Institute of Technology, Pasadena 91125, USA.
Wang
C M
CM
Garrity
P A
PA
Fraser
S E
SE
D000818:Animals; D004331:Drosophila melanogaster; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology; D008164:Luminescent Proteins / Q000032:analysis*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D009474:Neurons / Q000648:ultrastructure; D017785:Photons; D017956:Photoreceptor Cells, Invertebrate / Q000648:ultrastructure; D051381:Rats
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins