Abstract
We developed an optical oblique-incidence reflectivity difference (OI-RD) scanning microscope for imaging microarrays of label-free protein and DNA. Such a microscope complements currently widely used fluorescence-based optical microscopes by offering the capability to detect biochemical activities of DNA and protein molecules without the influence of fluorescent-labeling molecules. The specific activity and function of protein molecules are particularly subject to binding of small or large foreign molecules either directly through conformational change in the protein molecule itself or indirectly through properties of the attached molecules. We show that an OI-RD microscope can be used to: 1) detect binding reactions on microarrays without labeling, 2) quantitatively measure the optical properties of microarray spots, and 3) detect microarrays submerged in solution (which will enable OI-RD to monitor reaction kinetics on microarrays in reactive solutions). Furthermore, using both OI-RD and fluorescence images of an immunoglobulin-G (IgG) protein microarray, we observed that labeled and unlabeled IgG molecules deposited on the microarray substrate exhibit different wetting behaviors, and a mixture of the two tends to segregate into labeled and unlabeled regions. This illustrates potentially undesirable effects of fluorescent-labeling agents on protein properties that are of interest.
| Original language | English (US) |
|---|---|
| Title of host publication | Proceedings of SPIE - The International Society for Optical Engineering |
| Editors | D.V. Nicolau, R. Raghavachari |
| Pages | 121-128 |
| Number of pages | 8 |
| Volume | 5328 |
| DOIs | |
| State | Published - 2004 |
| Event | Microarrays and Combinatorial Techniques: Design, Fabrication, and Analysis II - San Jose, CA, United States Duration: Jan 25 2004 → Jan 26 2004 |
Other
| Other | Microarrays and Combinatorial Techniques: Design, Fabrication, and Analysis II |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 1/25/04 → 1/26/04 |
Fingerprint
Keywords
- Biochips
- Ellipsometry
- Hybridization
- Immunoglobulin-G
- In-situ detection
- Label-free detection
- Microarrays
- Oblique-incidence reflectivity difference
- OI-RD
- Oligonucleotide
ASJC Scopus subject areas
- Electrical and Electronic Engineering
- Condensed Matter Physics
Cite this
Detection of biomolecular microarrays without fluorescent-labeling agents. / Landry, J. P.; Zhu, X. D.; Gregg, Jeffrey; Guo, X. W.
Proceedings of SPIE - The International Society for Optical Engineering. ed. / D.V. Nicolau; R. Raghavachari. Vol. 5328 2004. p. 121-128.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Detection of biomolecular microarrays without fluorescent-labeling agents
AU - Landry, J. P.
AU - Zhu, X. D.
AU - Gregg, Jeffrey
AU - Guo, X. W.
PY - 2004
Y1 - 2004
N2 - We developed an optical oblique-incidence reflectivity difference (OI-RD) scanning microscope for imaging microarrays of label-free protein and DNA. Such a microscope complements currently widely used fluorescence-based optical microscopes by offering the capability to detect biochemical activities of DNA and protein molecules without the influence of fluorescent-labeling molecules. The specific activity and function of protein molecules are particularly subject to binding of small or large foreign molecules either directly through conformational change in the protein molecule itself or indirectly through properties of the attached molecules. We show that an OI-RD microscope can be used to: 1) detect binding reactions on microarrays without labeling, 2) quantitatively measure the optical properties of microarray spots, and 3) detect microarrays submerged in solution (which will enable OI-RD to monitor reaction kinetics on microarrays in reactive solutions). Furthermore, using both OI-RD and fluorescence images of an immunoglobulin-G (IgG) protein microarray, we observed that labeled and unlabeled IgG molecules deposited on the microarray substrate exhibit different wetting behaviors, and a mixture of the two tends to segregate into labeled and unlabeled regions. This illustrates potentially undesirable effects of fluorescent-labeling agents on protein properties that are of interest.
AB - We developed an optical oblique-incidence reflectivity difference (OI-RD) scanning microscope for imaging microarrays of label-free protein and DNA. Such a microscope complements currently widely used fluorescence-based optical microscopes by offering the capability to detect biochemical activities of DNA and protein molecules without the influence of fluorescent-labeling molecules. The specific activity and function of protein molecules are particularly subject to binding of small or large foreign molecules either directly through conformational change in the protein molecule itself or indirectly through properties of the attached molecules. We show that an OI-RD microscope can be used to: 1) detect binding reactions on microarrays without labeling, 2) quantitatively measure the optical properties of microarray spots, and 3) detect microarrays submerged in solution (which will enable OI-RD to monitor reaction kinetics on microarrays in reactive solutions). Furthermore, using both OI-RD and fluorescence images of an immunoglobulin-G (IgG) protein microarray, we observed that labeled and unlabeled IgG molecules deposited on the microarray substrate exhibit different wetting behaviors, and a mixture of the two tends to segregate into labeled and unlabeled regions. This illustrates potentially undesirable effects of fluorescent-labeling agents on protein properties that are of interest.
KW - Biochips
KW - Ellipsometry
KW - Hybridization
KW - Immunoglobulin-G
KW - In-situ detection
KW - Label-free detection
KW - Microarrays
KW - Oblique-incidence reflectivity difference
KW - OI-RD
KW - Oligonucleotide
UR - http://www.scopus.com/inward/record.url?scp=7644236346&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=7644236346&partnerID=8YFLogxK
U2 - 10.1117/12.524554
DO - 10.1117/12.524554
M3 - Conference contribution
VL - 5328
SP - 121
EP - 128
BT - Proceedings of SPIE - The International Society for Optical Engineering
A2 - Nicolau, D.V.
A2 - Raghavachari, R.
ER -