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001-es BibID:	BIBFORM057905
Első szerző:	Bene László (biofizikus)
Cím:	Dual-laser homo-FRET on the cell surface / László Bene, Tamás Ungvári, Roland Fedor, István Nagy, László Damjanovich
Dátum:	2015
ISSN:	0167-4889
Megjegyzések:	Inhomogeneous broadening and red-edge effects have been detected on a highly mobile system of fluorescently conjugated mAbs targeted to cell surface receptors. By exploiting site-selective spectroscopy and the characteristic loss of homo-FRET on increasing excitation and decreasing emission wavelengths, contributions of physical rotation and homo-FRET to the depolarization of fluorescence anisotropy have been separated. Absolute homo-FRET efficiency has been determined by ratioing two anisotropies: a homo-FRET-sensitive one, which is excited at the absorption main band and detected at the long wavelength region of emission, and a homo-FRET-insensitive one, which is excited at the long wavelength region of absorption and detected at the short wavelength region of emission. Because the anisotropies are simultaneously detected in a unified detection scheme of a dual T-format arrangement, the method is applicable for the real-time tracking of dynamical changes of physical rotations and proximities. The utility of the method is demonstrated in the context of the MHCII molecule and the heavy and light chains of the MHCI molecule, a system of three receptors with well-characterized close mutual proximities. Although the method is presented for a flow cytometer, it can also be realized in a fluorescence microscope capable for dual-laser excitation and dual-anisotropy detection.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Inhomogeneous broadening Red-edge effect Directed energy migration FRET (emFRET) Fluorescence anisotropy Fluorescence anisotropy lifetime imaging microscopy (rFLIM)
Megjelenés:	Biochimica et Biophysica Acta (BBA). Molecular Cell Research. - 1853 : 5 (2015), p. 1096-1112. -
További szerzők:	Ungvári Tamás Fedor Roland (1975-) (sebész) Nagy István (1957-) (villamosmérnök) Damjanovich László (1960-) (általános sebész)
Pályázati támogatás:	TÁMOP-4.2.2.A-11/1/KONV-2012-0045 TÁMOP Kardiológia Kutatócsoport OTKA Bridging Fund support OSTRAT/ 810/213 by the University of Debrecen OTKA
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