Label-Free and Real-Time Monitor of Binding and Dissociation Processes between Protein A and Swine IgG by Oblique-Incidence Reflectivity Difference Method

Life science has a need for detection methods that are label-free and real-time. In this paper, we have selected staphylococcal protein A （SPA） and swine immunoglobulin G （IgG）, and monitor the bindings between SPA and swine IgG with different concentrations, as well as the dissociations of SPA-swine IgG complex in different pH values of phosphate buffer by oblique-incidence reflectivity difference （OIRD） in a label-free and real-time fashion. We obtain the ON and OFF reaction dynamic curves corresponding to the bindings and dissociations of SPA and swine IgG. Through our analysis of the experimental results, we have been able to obtain the damping coefficients and the dissociation time of SPA and swine IgG for different pH values of the phosphate buffer. The results prove that the OIRD technique is a competing method for monitoring the dynamic processes of biomolecule interaction and achieving the quantitative information of reaction kinetics.

Label-free detection of hybridization of oligonucleotides by oblique-incidence reflectivity difference method

The microarrays of 20-base oligonucleotide with different concentrations are detected before and after hybridization by the oblique-incidence reflectivity difference (OI-RD) method. The experimental results prove that OI-RD is a label-free method which can not only distinguish the concentration difference of oligonucleotides before and after the hybridization but also detect the hybridization of short oligonucleotides. At present the OI-RD method can detect 0.39 μmol/L 20-base oligonucleotide or less. These results suggest that the OI-RD method is a promising and potential technique for label-free detection of biological microarrays.

Detection of hybridization of protein microarrays using an oblique-incidence reflectivity difference method

Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) method.Two detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained simultaneously.The detection results of hybridization by OI-RD were in accord with that of traditional fluorescent scans.In particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic coordinates.The detection sensitivity of OI-RD can reach 14 fg.These experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis.

Label-free detection repeatability of protein microarrays by oblique-incidence reflectivity difference method

We examine the repeatabilities of oblique-incidence reflectivity difference(OIRD) method for label-free detecting biological molecular interaction using protein microarrays.The experimental results show that the repeatabilities are the same in a given microarray or microarray-microarray and are consistent,indicating that OIRD is a promising label-free detection technique for biological microarrays.

Label-free and real-time detection of antigen-antibody interactions by Oblique-incidence Reflectivity Difference(OIRD) method

We label-free and real-time detected three interaction processes of antigen-antibodies,Human Immunoglobulin G(IgG),Rabbit IgG,and Mouse IgG as the targets,and Goat Anti-human IgG,Goat Anti-rabbit IgG,and Goat Anti-mouse IgG as the probe,by the Oblique-incidence Reflectivity Difference(OIRD) method.The interaction dynamic curves of the OIRD signal,corresponding to the interaction processes of antigen-antibodies,are generated.The reaction times from beginning to equilibrium state are about 1800,900,and 1200 s for Human IgG,Rabbit IgG,and Mouse IgG,respectively.The experimental results demonstrate that the OIRD method not only can distinguish biomolecular interactions,but also can be used in real-time detection of interactions and dynamic processes of biomolecules.

Label-free high-throughput and real-time detections of protein interactions by oblique-incidence reflectivity difference method

Selected Mouse IgG of 1 mg/mL as target was fabricated on microarray for 500 sample dots.Label-free and real-time reaction dynamic processes were detected between the microarrays with Goat Anti-mouse IgG of 0.02 mg/mL using the obliqueincidence reflectivity difference(OIRD)method.We obtained the reaction results and the reaction dynamic curves of 500 protein dots.In addition,we also used label-free detection of protein microarrays of 10080 sample dots,including BSA and different concentrations of Mouse IgG and Rabbit IgG,by OIRD.The obtained reaction results between the protein microarray with 1 mg/mL Goat Anti-mouse IgG and 1 mg/mL Goat Anti-rabbit IgG are reported herein.Experimental results show that OIRD can be not only label-free high-throughput detection method for biological microarrays but also label-free real-time detection in the interaction processes of biomolecules.

Experimental investigation of ghost imaging of reflective objects with different surface roughness

We present an experimental demonstration of ghost imaging of reflective objects with different surface roughness.The influence of the surface roughness, the transverse size of the test detector, and the reflective angle on the signal-to-noise ratio(SNR) is analyzed by measuring the second-order correlation of the light field based on classical statistical optics. It is shown that the SNR decreases with an increment of the surface roughness and the detector's transverse size or a decrease of the reflective angle. Additionally, the comparative studies between the rough object and the smooth one under the same conditions are also discussed.

New method for thickness determination and microscopic imaging of graphene-like two-dimensional materials

We employed the microscopic reflectance difference spectroscopy （micro-RDS） to determine the layer- number and microscopically image the surface topography of graphene and MoS2 samples. The contrast image shows the efficiency and reliability of this new clipping technique. As a low-cost, quantifiable, no-contact and non-destructive method, it is not concerned with the characteristic signal of certain materials and can be applied to arbitrary substrates. Therefore it is a perfect candidate for characterizing the thickness of graphene-like two- dimensional materials.

Giant in-plane optical anisotropy of a-plane ZnO on r-plane sapphire

We have measured the in-plane optical anisotropy （IPOA） of （1120） ZnO （a-plane） on （10]-2） sapphire （r-plane） by reflectance difference spectroscopy （RDS） at room temperature. Giant IPOA has been observed be- tween the light polarized direction parallel and perpendicular to the c axis of ZnO, since the symmetry of a-plane is C2v. A sharp resonance has been observed near the fundamental band gap, which is induced by the polarization- depend band gap shift. The sharp line shape is attributed to the exciton transition. The spectra fitting and differential spectra indicate the polarization-depend band energies. The giant IPOA is possible enhanced by anisotropy strain along and perpendicular to the c axis in the a-plane.