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Detection of functional groups and antibodies on microfabricated surfaces by confocal microscopy
Author(s) -
Nashat Amir H.,
Moronne Mario,
Ferrari Mauro
Publication year - 1998
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19981020)60:2<137::aid-bit1>3.0.co;2-o
Subject(s) - confocal , silanization , confocal microscopy , chemistry , fluorescein , fluorescence , covalent bond , fluorescence microscope , silanes , monolayer , biophysics , biochemistry , silane , organic chemistry , biology , physics , geometry , mathematics , quantum mechanics , microbiology and biotechnology
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 × 10 −5 fluors per nm 2 . © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 137–146, 1998.