Fluorescence lifetime imaging study of a thin protein layer on solid surfaces.
Understanding the fundamental interactions between proteins and solid surfaces is essential in the area of implantable medical devices. Fluorescence methods offer the sensitivity required to study the formation of the initial thin protein layers that mediate biocompatibility of materials. Thin protein layers (bovine serum albumin labelled with 1-anilino-8-naphthalenesulfonate, BSA-ANS) deposited on several surfaces (glass, silicon, stainless steel, polystyrene, and silver island film) were studied using confocal frequency domain Fluorescence Lifetime Imaging Microscopy (FLIM) and single-point multifrequency lifetime analysis techniques. FLIM provides spatial information about both fluorophores located on the surface and physicochemical parameters of the surface microenvironment. The average fluorescence lifetimes (tau(av)) of the adsorbed BSA-ANS generated by the contact between a protein solution and the material surface were measured by the multifrequency modulation and phase shift. Results indicate that tau(av) values of the albumin complexes on the surfaces (approximately 12 ns) are, in general, shorter than tau(av) found in the bulk solution (approximately 14 ns). For some surfaces, like polystyrene and silver island film the differences in tau(av) of the adsorbed BSA-ANS were found to be much greater. The differences in fluorescence lifetimes may indicate structural changes in the BSA protein induced by contact with the surface.
Togashi
Denisio M
DM
Nanoscale Biophotonics Laboratory, Department of Chemistry, National University of Ireland, Galway, Ireland. denisio.togashi@nuigalway.ie
Ryder
Alan G
AG
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D005898:Glass; D008856:Microscopy, Fluorescence; D011137:Polystyrenes; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012825:Silicon; D012834:Silver; D013232:Steel; D013499:Surface Properties
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D011137:Polystyrenes; D013232:Steel; D012710:Serum Albumin, Bovine; D012834:Silver; C027132:1-anilino-8-naphthalenesulfonate; D012825:Silicon