摘要:
Disclosed is a method to detect unlabeled nucleic acids (DNA and/or RNA) in a taxa, species, and organelle-specific fashion using surface plasmon resonance (SPR) imaging. Taxa-specific, species-specific, or organelle-specific nucleic acids are affixed to an SPR-suitable substrate. A nucleic acid sample to be analyzed is then contacted with the SPR-substrate and the substrate analyzed to determine the presence or absence of specific hybridization between the nucleic acids bound to the substrate and the nucleic acids contained in the sample. The method does not require that either the bound nucleic acids nor the sample nucleic acids be labeled. The method can be used to identify the source of nucleic acids, their sequence, as well as to identify organisms and place them within a given taxonomic hierarchy.
摘要:
Disclosed are methods for making surface plasmon resonance-capable arrays wherein molecules, such as proteins or nucleic acids, or cells, are adhered to a metal substrate. The metal substrates are modified by depositing an ω-modified alkanethiol monolayer to the substrate and then contacting the ω-modified monolayer with a heterobifunctional linking compound. Biomolecules or cells can then be attached to the heterobifunctional linking compound. Also disclosed are arrays wherein glutathione-containing molecules are immobilized on the substrate and GST-containing molecules are then specifically immobilized onto the substrate, taking advantage of the affinity between glutathione and GST.
摘要:
Disclosed is a process to construct multi-component biomolecule or cellular arrays suitable for use in SPR imaging studies of large molecule, cellular/molecular, and cell/cell interactions. Also disclosed are the resulting arrays. The success of the procedure hinges on the use of a reversible protecting group to modify reversibly ω-functionalized alkanethiols self-assembled on metal substrates. The arrays themselves include a metal substrate, a continuous layer of an identical ω-modified alkanthiol adhered to the metal substrate, and one or more discrete spots of biomolecules or cells directly bonded to the continuous layer of ω-modified alkenthiol. The areas of the continuous layer of ω-modified alkenthiol not covered by one of the discrete spots are covered by a background material resistant to non-specific protein binding.
摘要:
A surface plasmon resonance imaging apparatus provides an improved optical assembly allowing fixed source and detector operating with a horizontal test surface for a more compact design. In a preferred embodiment, a mechanical linkage of planar mirrors provides a single point adjustment of angle of incidence and angle of refraction while maintaining a constant optical axis of the source and detector.
摘要:
Disclosed is a process to construct multi-component biomolecule or cellular arrays suitable for use in SPR imaging studies of large molecule, cellular/molecular, and cell/cell interactions. The success of the procedure hinges on the use of a reversible protecting group to modify reversibly &ohgr;-functionalized alkanethiols self-assembled on metal substrates.
摘要:
SPR-compatible substrates for high density microarray fabrication and analyses are provided. Novel carbon-on-metal thin film substrate architecture permits the integration of surface plasmon resonance detection with photolithographically fabricated biomolecule arrays for the analysis of biomolecular interactions. The utility of the technology is shown in the analysis of specific DNA-DNA, DNA-RNA and DNA-protein binding interactions. These new substrates may be used to determine the secondary structure of RNA molecules, to probe the sequence-specific binding kinetics and affinity of proteins and small molecules, and as substrates for small-molecule combinatorial chemistry platforms for drug discovery applications.
摘要:
Disclosed is a process to construct multi-component biomolecule or cellular arrays suitable for use in SPR imaging studies of large molecule, cellular/molecular, and cell/cell interactions. The success of the procedure hinges on the use of a reversible protecting group to modify reversibly .omega.-functionalized alkanethiols self-assembled on metal substrates.
摘要:
Disclosed is a method for fabricating 1-dimensional micro-arrays using parallel micro-fluidic channels on chemically-modified metal, carbon, silicon, and/or germanium surfaces; a μL detection volume method that uses 2-dimensional nucleic acid micro-arrays formed by employing the 1-dimensional DNA micro-arrays in conjunction with a second set of parallel micro-fluidic channels for solution delivery, and the 1-dimensional and 2-dimensional arrays used in the methods. The methodology allows the rapid creation of 1- and 2-dimensional arrays for SPR imaging and fluorescence imaging of DNA-DNA, DNA-RNA, DNA-protein, and protein-protein binding events. The invention enables very small volumes necessary for a variety of bioassay applications to be analyzed by SPR. Target solution volumes as small as 200 pL can be analyzed.
摘要:
SPR-compatible substrates for high density microarray fabrication and analyses are provided. Novel carbon-on-metal thin film substrate architecture permits the integration of surface plasmon resonance detection with photolithographically fabricated biomolecule arrays for the analysis of biomolecular interactions. The utility of the technology is shown in the analysis of specific DNA-DNA, DNA-RNA and DNA-protein binding interactions. These new substrates may be used to determine the secondary structure of RNA molecules, to probe the sequence-specific binding kinetics and affinity of proteins and small molecules, and as substrates for small-molecule combinatorial chemistry platforms for drug discovery applications.
摘要:
A surface plasmon resonant device provides practical portable operation through the use of a low power high efficiency LED source and a high-efficiency prism sample cell pre-loaded with probe molecules and sealed for field use. A simple mechanical control allows adjustment of angulation of the light and camera for accurate response outside of the laboratory.