FRET microscopy

Basics, issues and advantages of FLIM-FRET imaging

Ammasi Periasamy, Nirmal Mazumder, Yuansheng Sun, Kathryn G. Christopher

    Research output: Contribution to journalArticle

    4 Citations (Scopus)

    Abstract

    Förster resonance energy transfer (FRET) is an effective and high resolution method to investigate protein–protein interaction in live or fixed specimens. The FRET technique is increasingly employed to evaluate the molecular mechanisms governing diverse cellular processes such as vesicular transport, signal transduction and the regulation of gene expression. For FRET to occur, protein moieties should be close together within 10 nm, the dipole moment of the fluorophore targeted to the proteins should have an appropriate orientation, and the spectral overlap of the donor emission with the acceptor absorption should be >30 %. FRET can be used to estimate the distance between interacting protein molecules in vivo or in vitro using light microscopy systems. Visible fluorescent proteins (VFPs) have been widely used as a FRET pair in addition to organic dyes. Light microscopy techniques including wide-field, confocal and multiphoton microscopy systems provide spatial information of the interacting proteins with nanometer resolution. For better interpretation and quantitation of the FRET signal the contaminations—also called spectral bleedthrough (SBT)—have to be removed.

    Original languageEnglish
    Pages (from-to)249-276
    Number of pages28
    JournalSpringer Series in Chemical Physics
    Volume111
    DOIs
    Publication statusPublished - 2015

    Fingerprint

    Energy transfer
    Microscopic examination
    energy transfer
    microscopy
    Imaging techniques
    proteins
    Proteins
    Optical microscopy
    Signal transduction
    Fluorophores
    gene expression
    Dipole moment
    Gene expression
    dipole moments
    Coloring Agents
    dyes
    Molecules
    high resolution
    estimates
    molecules

    All Science Journal Classification (ASJC) codes

    • Physical and Theoretical Chemistry

    Cite this

    Periasamy, Ammasi ; Mazumder, Nirmal ; Sun, Yuansheng ; Christopher, Kathryn G. / FRET microscopy : Basics, issues and advantages of FLIM-FRET imaging. In: Springer Series in Chemical Physics. 2015 ; Vol. 111. pp. 249-276.
    @article{2c099264d1ff499ea9de0dfc14ced10c,
    title = "FRET microscopy: Basics, issues and advantages of FLIM-FRET imaging",
    abstract = "F{\"o}rster resonance energy transfer (FRET) is an effective and high resolution method to investigate protein–protein interaction in live or fixed specimens. The FRET technique is increasingly employed to evaluate the molecular mechanisms governing diverse cellular processes such as vesicular transport, signal transduction and the regulation of gene expression. For FRET to occur, protein moieties should be close together within 10 nm, the dipole moment of the fluorophore targeted to the proteins should have an appropriate orientation, and the spectral overlap of the donor emission with the acceptor absorption should be >30 {\%}. FRET can be used to estimate the distance between interacting protein molecules in vivo or in vitro using light microscopy systems. Visible fluorescent proteins (VFPs) have been widely used as a FRET pair in addition to organic dyes. Light microscopy techniques including wide-field, confocal and multiphoton microscopy systems provide spatial information of the interacting proteins with nanometer resolution. For better interpretation and quantitation of the FRET signal the contaminations—also called spectral bleedthrough (SBT)—have to be removed.",
    author = "Ammasi Periasamy and Nirmal Mazumder and Yuansheng Sun and Christopher, {Kathryn G.}",
    year = "2015",
    doi = "10.1007/978-3-319-14929-5_7",
    language = "English",
    volume = "111",
    pages = "249--276",
    journal = "Springer Series in Chemical Physics",
    issn = "0172-6218",
    publisher = "Springer New York",

    }

    FRET microscopy : Basics, issues and advantages of FLIM-FRET imaging. / Periasamy, Ammasi; Mazumder, Nirmal; Sun, Yuansheng; Christopher, Kathryn G.

    In: Springer Series in Chemical Physics, Vol. 111, 2015, p. 249-276.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - FRET microscopy

    T2 - Basics, issues and advantages of FLIM-FRET imaging

    AU - Periasamy, Ammasi

    AU - Mazumder, Nirmal

    AU - Sun, Yuansheng

    AU - Christopher, Kathryn G.

    PY - 2015

    Y1 - 2015

    N2 - Förster resonance energy transfer (FRET) is an effective and high resolution method to investigate protein–protein interaction in live or fixed specimens. The FRET technique is increasingly employed to evaluate the molecular mechanisms governing diverse cellular processes such as vesicular transport, signal transduction and the regulation of gene expression. For FRET to occur, protein moieties should be close together within 10 nm, the dipole moment of the fluorophore targeted to the proteins should have an appropriate orientation, and the spectral overlap of the donor emission with the acceptor absorption should be >30 %. FRET can be used to estimate the distance between interacting protein molecules in vivo or in vitro using light microscopy systems. Visible fluorescent proteins (VFPs) have been widely used as a FRET pair in addition to organic dyes. Light microscopy techniques including wide-field, confocal and multiphoton microscopy systems provide spatial information of the interacting proteins with nanometer resolution. For better interpretation and quantitation of the FRET signal the contaminations—also called spectral bleedthrough (SBT)—have to be removed.

    AB - Förster resonance energy transfer (FRET) is an effective and high resolution method to investigate protein–protein interaction in live or fixed specimens. The FRET technique is increasingly employed to evaluate the molecular mechanisms governing diverse cellular processes such as vesicular transport, signal transduction and the regulation of gene expression. For FRET to occur, protein moieties should be close together within 10 nm, the dipole moment of the fluorophore targeted to the proteins should have an appropriate orientation, and the spectral overlap of the donor emission with the acceptor absorption should be >30 %. FRET can be used to estimate the distance between interacting protein molecules in vivo or in vitro using light microscopy systems. Visible fluorescent proteins (VFPs) have been widely used as a FRET pair in addition to organic dyes. Light microscopy techniques including wide-field, confocal and multiphoton microscopy systems provide spatial information of the interacting proteins with nanometer resolution. For better interpretation and quantitation of the FRET signal the contaminations—also called spectral bleedthrough (SBT)—have to be removed.

    UR - http://www.scopus.com/inward/record.url?scp=84927947150&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84927947150&partnerID=8YFLogxK

    U2 - 10.1007/978-3-319-14929-5_7

    DO - 10.1007/978-3-319-14929-5_7

    M3 - Article

    VL - 111

    SP - 249

    EP - 276

    JO - Springer Series in Chemical Physics

    JF - Springer Series in Chemical Physics

    SN - 0172-6218

    ER -