Detection of functional groups and antibodies on microfabricated surfaces by confocal microscopy

Amir H. Nashat, Mario Moronne, Mauro Ferrari

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

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 10-6 fluors per nm2.

Original languageEnglish (US)
Pages (from-to)137-146
Number of pages10
JournalBiotechnology and Bioengineering
Volume60
Issue number2
DOIs
StatePublished - Oct 20 1998

Keywords

  • Aminosilane
  • Antibody immobilization
  • Fluorescence confocal microscopy
  • Heterobifunctional crosslinker
  • Mercaptosilane
  • Microfabrication

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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