Three-dimensional localisation of fluorescence resonance energy transfer in living cells under two-photon excitation.
Three-dimensional (3-D) imaging of fluorescence resonance energy transfer (FRET) in human cells under two-photon excitation was demonstrated in this study. A sample was prepared by expressing a donor and an acceptor in living cells and using an antibody to secure the proximity of contact between the donor and the acceptor. The quenching of fluorescence emission of a donor in the double-labelled cells indicates the presence of FRET that occurred in these living cells. Because of the quadratic relation of the excitation power, 3-D localisation of FRET becomes possible.
Xu
M G
MG
Centre for Micro-Photonics, School of Biophysical Sciences and Electrical Engineering, Swinburne University of Technology, Hawthorn, Victoria, Australia.
Crimeen
B
B
Ludford-Menting
M J
MJ
Gan
X
X
Russell
S M
SM
Gu
M
M
D000818:Animals; D015703:Antigens, CD / Q000032:analysis / Q000378:metabolism; D002461:Cell Line, Transformed; D006224:Cricetinae; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000032:analysis / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D017785:Photons; D013050:Spectrometry, Fluorescence
D015703:Antigens, CD; D008164:Luminescent Proteins; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; C409504:blue fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins