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Development of a confocal optical system design for molecular imaging applications of biochip.

Huang G, Xu S, Zhu J, Deng C, Dong Z, Yang Y, Yang X, Wang X, Jin G - Int J Biomed Imaging (2007)

Bottom Line: A novel confocal optical system design and a dual laser confocal scanner have been developed to meet the requirements of highly sensitive detection of biomolecules on microarray chips, which is characterized by a long working distance (wd>3.0 mm), high numerical aperture (NA=0.72), and only 3 materials and 7 lenses used.This confocal optical system has a high scanning resolution, an excellent contrast and signal-to-noise ratio, and an efficiency of collected fluorescence of more than 2-fold better than that of other commercial confocal biochip scanners.Some applications of gene and protein imagings using the dual laser confocal scanner are described.

View Article: PubMed Central - PubMed

Affiliation: Medical Systems Biology Research Center, School of Medicine, Tsinghua University, Beijing 100084, China.

ABSTRACT
A novel confocal optical system design and a dual laser confocal scanner have been developed to meet the requirements of highly sensitive detection of biomolecules on microarray chips, which is characterized by a long working distance (wd>3.0 mm), high numerical aperture (NA=0.72), and only 3 materials and 7 lenses used. This confocal optical system has a high scanning resolution, an excellent contrast and signal-to-noise ratio, and an efficiency of collected fluorescence of more than 2-fold better than that of other commercial confocal biochip scanners. The scanner is as equally good for the molecular imaging detection of enclosed biochips as for the detection of biological samples on a slide surface covered with a cover-slip glass. Some applications of gene and protein imagings using the dual laser confocal scanner are described.

No MeSH data available.


The application of protein chip for autoantibodies in patients serum.
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Related In: Results  -  Collection


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fig9: The application of protein chip for autoantibodies in patients serum.

Mentions: The new confocal scanner is also useful for fluorescence detection of the immunoreactions of proteins on chips. We describe here a protein microarray chip for the parallel detection of autoantibodies in the serum of patients withautoimmune diseases, including systemic lupus erythmatosus (SLE), mixed connective tissue disease (MCTD), Sjögren's syndrome (SS), Sjögren's syndrome A (SSA), Sjögren's syndrome B(SSB), Smith (Sm), Ribonucleoprotein (RNP), Scleroderma (Scl), systemic sclerosis (SSc), dermotomyositis (DM), double-stranded DNA(dsDNA), Phosphate-Buffered Saline Tween-20(PBST), and polymyositis (PM). Purified autoantigens (SSA-52, SSA-60, SSB, Sm, RNP-68, Scl-70, Jo-1, dsDNA, centromere B, Ribosomal P0, and extracts of Hep-2 cells) are immobilized on the gel chip as shown in Figure 9(a), where QC—quality control, BC—blank control, RC—reaction control, NC—negative control, 1-Jo-1, 2-Sm, 3-Scl-70, 4-CENP-B, 5-dsDNA, 6-SSB, 7-SSA-52, 8-Extracts of Hep-2 cells, 9-SSA-60, 10-Ribosomal P0, 11-RNP-68. The protein microarray chip was incubated with 30L of a five diseases-mixed serum (diluted 1 : 100 with PBST) for 30 minutes at room temperature. After being rinsed and washed one time for 5 minutes with PBST, the chip was incubated with 30L of Cy3-labeled goat antihuman IgG antibody for 30 minutes at room temperature. After another rinse and 5-minutes PBST wash, the chip was briefly centrifuged to dry it. The mixed serum of SLE, SS, SSc, MCTD, DM, and PM positive sera patient was tested on the protein microarray chip, which was then scanned using our confocal scanner. The result of scanning image in Figure 9(b) clearly showed that this mixed sera contained anti-Jo-1 at row 2 from column 1 to column 3, anti-Sm at row 2 from column 4 to column 6, anti-Scl-70 at row 2 from column 7 to column 9, anticentromere B at row 3 from column 1 to column 3, anti-dsDNA at row 3 from column 4 to column 6, anti-SSB at row 3 from column 7 to column 9, anti-SSA-52 at row 4 from column 1 to column 3, antinuclear antibodies at row 4 from column 7 to column 9, anti-SSA-60 at row 5 from column 1 to column 3, anti-Ribosomal P0 at row 5 from column 4 to column 6, and anti-RNP-68 at row 5 from column 7 to column 9.


Development of a confocal optical system design for molecular imaging applications of biochip.

Huang G, Xu S, Zhu J, Deng C, Dong Z, Yang Y, Yang X, Wang X, Jin G - Int J Biomed Imaging (2007)

The application of protein chip for autoantibodies in patients serum.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211516&req=5

fig9: The application of protein chip for autoantibodies in patients serum.
Mentions: The new confocal scanner is also useful for fluorescence detection of the immunoreactions of proteins on chips. We describe here a protein microarray chip for the parallel detection of autoantibodies in the serum of patients withautoimmune diseases, including systemic lupus erythmatosus (SLE), mixed connective tissue disease (MCTD), Sjögren's syndrome (SS), Sjögren's syndrome A (SSA), Sjögren's syndrome B(SSB), Smith (Sm), Ribonucleoprotein (RNP), Scleroderma (Scl), systemic sclerosis (SSc), dermotomyositis (DM), double-stranded DNA(dsDNA), Phosphate-Buffered Saline Tween-20(PBST), and polymyositis (PM). Purified autoantigens (SSA-52, SSA-60, SSB, Sm, RNP-68, Scl-70, Jo-1, dsDNA, centromere B, Ribosomal P0, and extracts of Hep-2 cells) are immobilized on the gel chip as shown in Figure 9(a), where QC—quality control, BC—blank control, RC—reaction control, NC—negative control, 1-Jo-1, 2-Sm, 3-Scl-70, 4-CENP-B, 5-dsDNA, 6-SSB, 7-SSA-52, 8-Extracts of Hep-2 cells, 9-SSA-60, 10-Ribosomal P0, 11-RNP-68. The protein microarray chip was incubated with 30L of a five diseases-mixed serum (diluted 1 : 100 with PBST) for 30 minutes at room temperature. After being rinsed and washed one time for 5 minutes with PBST, the chip was incubated with 30L of Cy3-labeled goat antihuman IgG antibody for 30 minutes at room temperature. After another rinse and 5-minutes PBST wash, the chip was briefly centrifuged to dry it. The mixed serum of SLE, SS, SSc, MCTD, DM, and PM positive sera patient was tested on the protein microarray chip, which was then scanned using our confocal scanner. The result of scanning image in Figure 9(b) clearly showed that this mixed sera contained anti-Jo-1 at row 2 from column 1 to column 3, anti-Sm at row 2 from column 4 to column 6, anti-Scl-70 at row 2 from column 7 to column 9, anticentromere B at row 3 from column 1 to column 3, anti-dsDNA at row 3 from column 4 to column 6, anti-SSB at row 3 from column 7 to column 9, anti-SSA-52 at row 4 from column 1 to column 3, antinuclear antibodies at row 4 from column 7 to column 9, anti-SSA-60 at row 5 from column 1 to column 3, anti-Ribosomal P0 at row 5 from column 4 to column 6, and anti-RNP-68 at row 5 from column 7 to column 9.

Bottom Line: A novel confocal optical system design and a dual laser confocal scanner have been developed to meet the requirements of highly sensitive detection of biomolecules on microarray chips, which is characterized by a long working distance (wd>3.0 mm), high numerical aperture (NA=0.72), and only 3 materials and 7 lenses used.This confocal optical system has a high scanning resolution, an excellent contrast and signal-to-noise ratio, and an efficiency of collected fluorescence of more than 2-fold better than that of other commercial confocal biochip scanners.Some applications of gene and protein imagings using the dual laser confocal scanner are described.

View Article: PubMed Central - PubMed

Affiliation: Medical Systems Biology Research Center, School of Medicine, Tsinghua University, Beijing 100084, China.

ABSTRACT
A novel confocal optical system design and a dual laser confocal scanner have been developed to meet the requirements of highly sensitive detection of biomolecules on microarray chips, which is characterized by a long working distance (wd>3.0 mm), high numerical aperture (NA=0.72), and only 3 materials and 7 lenses used. This confocal optical system has a high scanning resolution, an excellent contrast and signal-to-noise ratio, and an efficiency of collected fluorescence of more than 2-fold better than that of other commercial confocal biochip scanners. The scanner is as equally good for the molecular imaging detection of enclosed biochips as for the detection of biological samples on a slide surface covered with a cover-slip glass. Some applications of gene and protein imagings using the dual laser confocal scanner are described.

No MeSH data available.