Abstract
We describe a new oblique-incidence reflectivity difference (OI-RD) scanning microscope for high-throughput screening, in microarray format on functionalized glass slides, small-molecule compound libraries for protein ligands. The microscope employs a combination of scan mirror for y-scan and single-axis translation stage for x-scan. For a printed microarray with over 10,000 features, each of 100 ?m in diameter and distinct small molecule targets, we can acquire an end-point image of the microarray in 90 minutes with pixel resolution of 20 μm × 20μm. The microscope is also capable of measuring binding kinetics of over 10,000 protein-ligand reactions simultaneously. We also describe a number of strategies for immobilizing small molecule compounds on functionalized glass slides: (1) conjugating the compounds (through a chemically inert linker) with a lysine residue so that the primary amine on the lysine serves as the anchor to epoxy-functionalized glass surface; (2) conjugating the compounds (through a linker) with a biotin residue so that the biotin serves as the anchor to streptavidin-functionalized glass surface; (3) immobilizing small molecule compounds without modification on isocyanate-functionalized glass surface through non-specific reaction of nucleophilic molecular motifs on most bioactive compounds with isocyanate groups. We present preliminary measurements of protein-small molecule binding reactions using the new microscope and the surface immobilization strategies.
| Original language | English (US) |
|---|---|
| Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Volume | 7182 |
| DOIs | |
| State | Published - 2009 |
| Event | Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII - San Jose, CA, United States Duration: Jan 26 2009 → Jan 28 2009 |
Other
| Other | Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/26/09 → 1/28/09 |
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Keywords
- High-throughput
- Immobilization strategies
- In situ detection
- Label-free detection
- Microarrays
- Oblique-incidence reflectivity difference
- OI-RD
ASJC Scopus subject areas
- Applied Mathematics
- Computer Science Applications
- Electrical and Electronic Engineering
- Electronic, Optical and Magnetic Materials
- Condensed Matter Physics
Cite this
Label-free screening small-molecule compound libraries for protein-ligands using a high-throughput optical scanning microscope. / Fei, Y. Y.; Landry, J. P.; Sun, Y. S.; Zhu, X. D.; Wang, X. B.; Luo, J. T.; Wu, C. Y.; Lam, Kit.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7182 2009. 71820Q.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Label-free screening small-molecule compound libraries for protein-ligands using a high-throughput optical scanning microscope
AU - Fei, Y. Y.
AU - Landry, J. P.
AU - Sun, Y. S.
AU - Zhu, X. D.
AU - Wang, X. B.
AU - Luo, J. T.
AU - Wu, C. Y.
AU - Lam, Kit
PY - 2009
Y1 - 2009
N2 - We describe a new oblique-incidence reflectivity difference (OI-RD) scanning microscope for high-throughput screening, in microarray format on functionalized glass slides, small-molecule compound libraries for protein ligands. The microscope employs a combination of scan mirror for y-scan and single-axis translation stage for x-scan. For a printed microarray with over 10,000 features, each of 100 ?m in diameter and distinct small molecule targets, we can acquire an end-point image of the microarray in 90 minutes with pixel resolution of 20 μm × 20μm. The microscope is also capable of measuring binding kinetics of over 10,000 protein-ligand reactions simultaneously. We also describe a number of strategies for immobilizing small molecule compounds on functionalized glass slides: (1) conjugating the compounds (through a chemically inert linker) with a lysine residue so that the primary amine on the lysine serves as the anchor to epoxy-functionalized glass surface; (2) conjugating the compounds (through a linker) with a biotin residue so that the biotin serves as the anchor to streptavidin-functionalized glass surface; (3) immobilizing small molecule compounds without modification on isocyanate-functionalized glass surface through non-specific reaction of nucleophilic molecular motifs on most bioactive compounds with isocyanate groups. We present preliminary measurements of protein-small molecule binding reactions using the new microscope and the surface immobilization strategies.
AB - We describe a new oblique-incidence reflectivity difference (OI-RD) scanning microscope for high-throughput screening, in microarray format on functionalized glass slides, small-molecule compound libraries for protein ligands. The microscope employs a combination of scan mirror for y-scan and single-axis translation stage for x-scan. For a printed microarray with over 10,000 features, each of 100 ?m in diameter and distinct small molecule targets, we can acquire an end-point image of the microarray in 90 minutes with pixel resolution of 20 μm × 20μm. The microscope is also capable of measuring binding kinetics of over 10,000 protein-ligand reactions simultaneously. We also describe a number of strategies for immobilizing small molecule compounds on functionalized glass slides: (1) conjugating the compounds (through a chemically inert linker) with a lysine residue so that the primary amine on the lysine serves as the anchor to epoxy-functionalized glass surface; (2) conjugating the compounds (through a linker) with a biotin residue so that the biotin serves as the anchor to streptavidin-functionalized glass surface; (3) immobilizing small molecule compounds without modification on isocyanate-functionalized glass surface through non-specific reaction of nucleophilic molecular motifs on most bioactive compounds with isocyanate groups. We present preliminary measurements of protein-small molecule binding reactions using the new microscope and the surface immobilization strategies.
KW - High-throughput
KW - Immobilization strategies
KW - In situ detection
KW - Label-free detection
KW - Microarrays
KW - Oblique-incidence reflectivity difference
KW - OI-RD
UR - http://www.scopus.com/inward/record.url?scp=65949088815&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=65949088815&partnerID=8YFLogxK
U2 - 10.1117/12.809700
DO - 10.1117/12.809700
M3 - Conference contribution
VL - 7182
BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
ER -