title: Biosensor for direct cell detection, quantification and analysis.
abstract: Microarrays are promising tools for cell isolation and detection. However, they have yet to be widely applied in biology. This stems from a lack of demonstration of their sensitivity and compatibility with complex biological samples, and a lack of proof that their use does not induce aberrant cellular effects. Herein, we characterized and optimized a recently developed technology associating antibody microarrays with surface plasmon resonance imaging (SPRi). Using a murine macrophage cell line we demonstrate the binding specificity of our antibody-microarrays and the correlation between SPRi signals and both the number of bound cells, and the level of expression of cell surface markers. Confocal microscopy reveals that cell binding to the chip through antibody-antigen interactions underwent morphological changes reflecting the density of the relevant cell surface marker without affecting cell viability as shown by fluorescent microscopy. The detection threshold of the microarray-SPRi system is lowered 10-fold by applying a polyethylene oxide film to the gold surface of the chip. This increased sensitivity allows the detection of cells representing as little as 0.5% of a mixed population. The potential of this method is illustrated by two applications: characterization of ligand-cell receptor interactions, allowing determination of receptor specificity, and analysis of peripheral blood mononuclear cells, demonstrating the suitability of this tool for the analysis of complex biological samples.
authors.affiliation: INSERM, U823, Grenoble, F-38042, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Cortès
authors.forename: Christian L
authors.initials: CL
authors.lastname: Villiers
authors.forename: Pascal
authors.initials: P
authors.lastname: Colpo
authors.forename: Rachel
authors.initials: R
authors.lastname: Couderc
authors.forename: Carine
authors.initials: C
authors.lastname: Brakha
authors.forename: François
authors.initials: F
authors.lastname: Rossi
authors.forename: Patrice N
authors.initials: PN
authors.lastname: Marche
authors.forename: Marie-Bernadette
authors.initials: MB
authors.lastname: Villiers
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques / Q000379:methods* / Q000706:statistics & numerical data; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods* / Q000706:statistics & numerical data; D018922:HL-60 Cells; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D051379:Mice; D046228:Microarray Analysis; D018613:Microscopy, Confocal; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D055910:Antibodies, Immobilized; D011956:Receptors, Cell Surface