1 July 2003 Spectroscopic approach for monitoring two-photon excited fluorescence resonance energy transfer from homodimers at the subcellular level
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J. of Biomedical Optics, 8(3), (2003). doi:10.1117/1.1584052
Abstract
We have employed a spectroscopic approach for monitoring fluorescence resonance energy transfer (FRET) in living cells. This method provides excellent spectral separation of green fluorescent protein (GFP) mutant signals within a subcellular imaging volume using two-photon excited fluorescence imaging and spectroscopy (TPIS-FRET). In contrast to current FRET-based methodologies, TPIS-FRET does not rely on the selection of optical filters, ratiometric image analysis, or bleedthrough correction algorithms. Utilizing the intrinsic optical sectioning capabilities of TPIS-FRET, we have identified protein–protein interactions within discrete subcellular domains. To illustrate the applicability of this technique to the detection of homodimer formation, we demonstrated the in vivo association of promyleocyte (PML) homodimers within their corresponding nuclear body.
Vickie J. LaMorte, Aikaterini Zoumi, Bruce J. Tromberg, "Spectroscopic approach for monitoring two-photon excited fluorescence resonance energy transfer from homodimers at the subcellular level," Journal of Biomedical Optics 8(3), (1 July 2003). http://dx.doi.org/10.1117/1.1584052
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KEYWORDS
Fluorescence resonance energy transfer

Proteins

Luminescence

Green fluorescent protein

Spectroscopy

Optical filters

Microscopes

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