Open AccessOptical properties of Alexa™ 488 and Cy™5 immobilized on a glass surface
Author(s) -
Lili Wang,
Adolfas K. Gaigalas,
Vytas Reipa
Publication year - 2005
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/05381mt03
Subject(s) - fluorophore , fluorescence , absorption (acoustics) , analytical chemistry (journal) , ellipsometry , chemistry , aqueous solution , alexa fluor , polarizability , emission spectrum , fluorescence spectroscopy , absorption spectroscopy , spectral line , materials science , photochemistry , optics , chromatography , nanotechnology , molecule , thin film , organic chemistry , astronomy , composite material , physics
The absorption and emission spectra were measured for Cy™5 and Alexa™ 488 fluorophores confined on a glass surface. The data were obtained using fluorometry and spectroscopic ellipsometry. Red shifts of the surface-immobilized fluorophore absorption spectra relative to the fluorophore spectra in aqueous solution were observed using both methods. We interpret these red shifts in terms of a change in the polarizability and polarity of the effective solvent. A formula is given that can be used to estimate expected shifts in absorption and emission maxima for surface-immobilized fluorophores. Spectroscopic ellipsometry measurements provide identification of the fluorophores confined on a glass surface. These results suggest that the design of microarray detection systems should be based on the optical properties of fluorophores attached to the surface and not on the optical properties of fluorophores in solution.