Detection of functional groups and antibodies on microfabricated surfaces by confocal microscopy.
Fluorescence confocal microscopy was used to characterize micron-sized microfabricated silicon particles and planar oxide surfaces after silanization and immobilization of IgG antibody. Surfaces treated with amino- and mercaptosilanes were tested for the presence of amine and sulfhydryl groups by labeling with specific fluorescein probes. In addition, human antibody (IgG) was immobilized to the thiol-coated microparticles using the heterobifunctional crosslinker succinimidyl 4-(N-maleimidolmethyl)-cyclohexane-1-carboxylate. Estimates of the surface density of IgG were consistent with 8.3% of a monolayer of covalently-bound antibody. Confocal images confirmed uniform layers of both silanes and antibodies on the microparticles. The sensitivity limit for the confocal measurements was determined to be as low as 1.5 x 10(-5) fluors per nm2.
Nashat
A H
AH
Department of Materials Science and Mineral Engineering, University of California at Berkeley, Berkeley, California 94720, USA.
Moronne
M
M
Ferrari
M
M
D000704:Analysis of Variance; D003432:Cross-Linking Reagents; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D008301:Maleimides; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D010087:Oxides; D012821:Silanes; D012825:Silicon; D013499:Surface Properties
D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D008301:Maleimides; D010087:Oxides; D012821:Silanes; C025285:N-(4-carboxycyclohexylmethyl)maleimide N-hydroxysuccinimide ester; D016650:Fluorescein-5-isothiocyanate; D012825:Silicon