Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling

Jonathan S. Marchant, Grace E. Stutzmann, Malcolm A. Leissring, Frank M. LaFerla, Ian Parker

Research output: Contribution to journalArticle

87 Citations (Scopus)

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 (λ <760 nm) rapidly changes the fluorescence of DsRed from red to green when viewed subsequently by conventional (one-photon) epifluorescence. Mechanistically, three-photon excitation (λ <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.

Original languageEnglish (US)
Pages (from-to)645-649
Number of pages5
JournalNature Biotechnology
Volume19
Issue number7
DOIs
StatePublished - 2001
Externally publishedYes

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Organelles
Labeling
Fusion reactions
Color
Proteins
Cell Fusion
Cell Lineage
Protein Transport
Photons
Cell Survival
Cells
Imaging techniques
Population
fluorescent protein 583
red fluorescent protein

ASJC Scopus subject areas

  • Microbiology

Cite this

Marchant, J. S., Stutzmann, G. E., Leissring, M. A., LaFerla, F. M., & Parker, I. (2001). Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling. Nature Biotechnology, 19(7), 645-649. https://doi.org/10.1038/90249

Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling. / Marchant, Jonathan S.; Stutzmann, Grace E.; Leissring, Malcolm A.; LaFerla, Frank M.; Parker, Ian.

In: Nature Biotechnology, Vol. 19, No. 7, 2001, p. 645-649.

Research output: Contribution to journalArticle

Marchant, JS, Stutzmann, GE, Leissring, MA, LaFerla, FM & Parker, I 2001, 'Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling', Nature Biotechnology, vol. 19, no. 7, pp. 645-649. https://doi.org/10.1038/90249
Marchant, Jonathan S. ; Stutzmann, Grace E. ; Leissring, Malcolm A. ; LaFerla, Frank M. ; Parker, Ian. / Multiphoton-evoked color change of DsRed as an optical highlighter for cellular and subcellular labeling. In: Nature Biotechnology. 2001 ; Vol. 19, No. 7. pp. 645-649.
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