A novel high-throughput scanning microscope for label-free detection of protein and small-molecule chemical microarrays.

We describe a novel scanning optical microscope based on a polarization-modulated nulling ellipsometry. The new microscope employs a combination of scanning mirror and sample translation and thus enables high-throughput label-free detection of biomolecular microarrays with more than 10 000 protein or small-molecule targets. For illustration, we show the image of a 2760-spot protein microarray on a functionalized glass slide obtained with such a microscope. The new scanning microscope is also capable of determining, in parallel, the real-time binding kinetics of multiple molecular species under aqueous conditions.

[1]  G. Scatchard,et al.  Physical-chemical characteristics of certain of the proteins of normal human plasma. , 1947, The Journal of physical and colloid chemistry.

[2]  M R Thompson,et al.  Hydrodynamic structure of bovine serum albumin determined by transient electric birefringence. , 1975, Biophysical journal.

[3]  R. Azzam,et al.  Ellipsometry and polarized light : North Holland, Amsterdam, 1987 (ISBN 0-444-87016-4). xvii + 539 pp. Price Dfl. 75.00. , 1987 .

[4]  Hans Arwin,et al.  Imaging ellipsometry revisited: Developments for visualization of thin transparent layers on silicon substrates , 1996 .

[5]  J. Weisel,et al.  Novel Interactions between Urokinase and Its Receptor* , 2000, The Journal of Biological Chemistry.

[6]  T. Kodadek Protein microarrays: prospects and problems. , 2001, Chemistry & biology.

[7]  P. Mitchell A perspective on protein microarrays , 2002, Nature Biotechnology.

[8]  Gavin MacBeath,et al.  Protein microarrays and proteomics , 2002, Nature Genetics.

[9]  G. Jin,et al.  A label-free multisensing immunosensor based on imaging ellipsometry. , 2003, Analytical chemistry.

[10]  R. Karlsson,et al.  SPR for molecular interaction analysis: a review of emerging application areas , 2004, Journal of molecular recognition : JMR.

[11]  Kit S. Lam,et al.  A novel approach to chemical microarray using ketone-modified macromolecular scaffolds: Application in micro cell-adhesion assay , 2004, Molecular Diversity.

[12]  J P Landry,et al.  Label-free detection of microarrays of biomolecules by oblique-incidence reflectivity difference microscopy. , 2004, Optics letters.

[13]  C. Y. Fong,et al.  An oblique-incidence optical reflectivity difference and LEED study of rare-gas growth on a lattice-mismatched metal substrate , 2004 .

[14]  Charles T Campbell,et al.  Quantitative methods for spatially resolved adsorption/desorption measurements in real time by surface plasmon resonance microscopy. , 2004, Analytical chemistry.

[15]  H. Koga,et al.  A novel approach to protein expression profiling using antibody microarrays combined with surface plasmon resonance technology , 2005, Proteomics.

[16]  Timothy Londergan,et al.  Looking towards label-free biomolecular interaction analysis in a high-throughput format: a review of new surface plasmon resonance technologies. , 2006, Current opinion in biotechnology.

[17]  A. Hillier,et al.  Surface plasmon resonance imaging of biomolecular interactions on a grating-based sensor array. , 2006, Analytical chemistry.

[18]  Kit S. Lam,et al.  Oblique-incidence reflectivity difference microscope for label-free high-throughput detection of biochemical reactions in a microarray format. , 2007 .