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001-es BibID:	BIBFORM056493
Első szerző:	Bene László (biofizikus)
Cím:	Single-laser polarization FRET (polFRET) on the cell surface / László Bene, Tamás Ungvári, Roland Fedor, László Damjanovich
Dátum:	2014
ISSN:	0167-4889
Megjegyzések:	A new method for the simultaneous detection of rotational mobility and proximity of cell surface receptors is presented based on cell-by-cell basis measurement of polarized fluorescence intensity components of the donor and acceptor of a FRET system. In addition to the FRET efficiency and the donor and acceptor concentrations, the method makes also possible the determination of the rotational characteristics and the associated fraction of the donors (FRET-fraction). The method is illustrated with flow cytometric and rFLIM measurements on donor-acceptor systems comprising fluorescently labeled whole antibodies and their Fab fragments against epitopes of the MHCI and MHCII cell surface receptors on human lymphoblast cells. Fluorescence anisotropy of donor and acceptor and FRET efficiency were measured for samples of different acceptor-to-donor concentration ratios. Acceptor anisotropy proved to be more sensitive than the donor anisotropy for sensing FRET. After determining the rotational constants of the donor-conjugated antibodies by measurements of FRET in the steady state, and by rFLIM as a reference, the associated fractions of the MHCI and MHCII molecules in their clusters were determined. Besides the flow cytometer and the wide-field rFLIM used in this study, the method can be applied also in other devices capable of dual-anisotropy detection.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Fluorescence anisotropy Rotational mobility Proximity Receptor cluster FRET-fraction rFLIM
Megjelenés:	Biochimica et Biophysica Acta (BBA). Molecular Cell Research. - 1843 : 12 (2014), p. 3047-3064. -
További szerzők:	Ungvári Tamás Fedor Roland (1975-) (sebész) Damjanovich László (1960-) (általános sebész)
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat DOI Szerző által megadott URL
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