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Land-contrast self-referencing interferometric protein microarray
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View: Figures


Image of FIG. 1.
FIG. 1.

The principle of the LC BioCD (in chip format). (a1) Spot patterns were etched on silicon thermal oxide chips by photolithography (140 nm on spots/77 nm on land). (a2) The spot and land have the same reflectance at 633 nm wavelength (normal incidence). The spot-land contrast is zero. Black circles in the picture are due to light scattering on the edge of the spots. (a3) The spot-land contrast appears after evenly applying an antibody layer on the entire chip. (a4) The contrast continues to increase after a sandwich assay. (b) LC chips work in both air and water environment, and the working condition 77 nm/140 nm is suitable for both cases. (c) The calculated reflectance changes to a protein layer are opposite and optimized on the spot and land. The contrast-protein conversion factor is calculated to be 2.55% per 1 nm protein in air and 0.51% in water.

Image of FIG. 2.
FIG. 2.

The kinetic curves of rabbit IgG forward and sandwich assays. (a) The complete process of the immunoassays (including antibody immobilization, forward assay, and sandwich assay) at five concentrations monitored in real time. Flow rate is . 1000 ng/ml mouse IgG and 0 ng/ml rabbit IgG are controls. (b) The forward assay part of (a) was analyzed and fitted by the real-time binding equation [Eq. (5)]. The relevant binding parameters , , , and maximum antigen layer height were calculated from the and fits.

Image of FIG. 3.
FIG. 3.

The response curve of rabbit IgG sandwich assay read in the dry state. antirabbit IgG in PBS was incubated on eight wells of a LC BioCD for 30 min to form an antibody layer. Concentrations of 0, 10, 30, 100, 300, 1000, 3000, and rabbit IgG were incubated on the eight wells for 30 min, and then incubated under antirabbit secondary on all wells for 30 min. The contrast differences before and after the sandwich are shown for 30, 100, 300, 1000, and 3000 ng/ml concentration groups.


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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Land-contrast self-referencing interferometric protein microarray