Enzyme-containing hydrogel micropatterns serving a dual purpose of cell sequestration and metabolite detection

Jun Yan, Yinghua Sun, He Zhu, Laura Marcu, Alexander Revzin

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

The integration of sensing elements with small groups of cells is a critical step towards miniaturization of cell cultivation and analysis. This paper describes the development of an optical, enzyme-based biosensor for local detection of hydrogen peroxide (H2O2) secreted by stimulated macrophages. Photolithographic patterning of horseradish peroxidase (HRP)-containing poly (ethylene glycol) (PEG) hydrogel microstructures was used to create sensing structures on the glass surface. Importantly, enzyme-entrapping hydrogel micropatterns did not support protein or cell deposition and allowed to guide attachment of macrophages next to the sensing elements. Amplex Red, an organic molecule that becomes fluorescent in the presence of H2O2 and HRP, was either immobilized inside hydrogel elements alongside enzyme molecules or added into the cell culture media during cell activation. The production of H2O2 after mitogenic stimulation of macrophages resulted in appearance of fluorescence in the HRP-containing hydrogel microstructures, with fluorescence intensity being a strong function of analyte concentration. The novel cell culture system with integrated sensing elements described here may be enhanced in the future by incorporating additional biorecognition elements to enable multi-metabolite detection at the site of a cell.

Original languageEnglish (US)
Pages (from-to)2604-2610
Number of pages7
JournalBiosensors and Bioelectronics
Volume24
Issue number8
DOIs
StatePublished - Apr 15 2009

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Keywords

  • Biosensors of extracellular metabolites
  • Cell micropatterning
  • Hyrogel microstructures

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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