A novel method for observing proteins in vivo using a small fluorescent label and multiphoton imaging.
A novel method for the fluorescence detection of proteins in cells is described in the present study. Proteins are labelled by the selective biosynthetic incorporation of 5-hydroxytryptophan and the label is detected via selective two-photon excitation of the hydroxyindole and detection of its fluorescence emission at 340 nm. The method is demonstrated in this paper with images of a labelled protein in yeast cells.
Botchway
Stanley W
SW
Central Laser Facility, Rutherford Appleton Laboratory, Chilton, Didcot, Oxfordshire OX11 0QX, UK.
Barba
Ignasi
I
Jordan
Randolf
R
Harmston
Rebecca
R
Haggie
Peter M
PM
Williams
Simon-Peter
SP
Fulton
Alexandra M
AM
Parker
Anthony W
AW
Brindle
Kevin M
KM
D006916:5-Hydroxytryptophan / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis*; D008856:Microscopy, Fluorescence; D010735:Phosphoglycerate Kinase / Q000737:chemistry / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D006916:5-Hydroxytryptophan; D010735:Phosphoglycerate Kinase