Faculty Collaboration Database

Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling. Nat Biotechnol 2001 Jul;19(7):645-9

Date

07/04/2001

Pubmed ID

11433276

DOI

10.1038/90249

Scopus ID

2-s2.0-0034930509 (requires institutional sign-in at Scopus site)   93 Citations

Abstract

DsRed, a recently cloned red fluorescent protein, has attracted great interest as an expression tracer and fusion partner for multicolor imaging. We report that three-photon excitation (lambda <760 nm) rapidly changes the fluorescence of DsRed from red to green when viewed subsequently by conventional (one-photon) epifluorescence. Mechanistically, three-photon excitation (lambda <760 nm) selectively bleaches the mature, red-emitting form of DsRed, thereby enhancing emission from the immature green form through reduction of fluorescence resonance energy transfer (FRET). The "greening" effect occurs in live mammalian cells at the cellular and subcellular levels, and the resultant color change persists for >30 h without affecting cell viability. This technique allows individual cells, organelles, and fusion proteins to be optically marked and has potential utility for studying cell lineage, organelle dynamics, and protein trafficking, as well as for selective retrieval of cells from a population. We describe optimal parameters to induce the color change of DsRed, and demonstrate applications that show the potential of this optical highlighter.

Author List

Marchant JS, Stutzmann GE, Leissring MA, LaFerla FM, Parker I

Author

Jonathan S. Marchant PhD Chair, Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

3T3 Cells
Animals
CHO Cells
Cell Line
Cricetinae
Fluorescent Dyes
Humans
Luminescent Proteins
Mice
Microscopy, Confocal
Microscopy, Fluorescence
Photons
Plasmids
Recombinant Fusion Proteins
Time Factors