The present invention pertains to the field of pre-functionalised nanoparticles (NPs). It relates more particularly to NPs pre functionalized using a self-assembled monolayer (SAM) and also to NPs functionalized using biomolecules such that the NPs are stable in solution. These NPs may be used in numerous applications, especially as a diagnostic tool, tool for depleting a molecule of interest in a solution, and therapeutic tool.

The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.

In an example method, a hydrogel is applied to a surface of a substrate and primers are grafted to the applied hydrogel. Before or after the primers are grafted, plasmonic nanostructures are introduced to the applied hydrogel. This substrate can make up one surface of a flow cell. When the flow cell is used in a sequencing operation, the plasmonic nanostructures can enhance fluorescent signals that are generated.

In an example method, a hydrogel is applied to a surface of a substrate and primers are grafted to the applied hydrogel. Before or after the primers are grafted, plasmonic nanostructures are introduced to the applied hydrogel. This substrate can make up one surface of a flow cell. When the flow cell is used in a sequencing operation, the plasmonic nanostructures can enhance fluorescent signals that are generated.

An example of an incorporation mix includes a liquid carrier, a complex, and a labeled nucleotide. The complex includes a polymerase and a plasmonic nanostructure linked to the polymerase. The labeled nucleotide includes a nucleotide, a 3′ OH blocking group attached to a sugar of the nucleotide, and a dye label attached to a base of the nucleotide.

An example of an incorporation mix includes a liquid carrier, a complex, and a labeled nucleotide. The complex includes a polymerase and a plasmonic nanostructure linked to the polymerase. The labeled nucleotide includes a nucleotide, a 3′ OH blocking group attached to a sugar of the nucleotide, and a dye label attached to a base of the nucleotide.

An example of a functionalized plasmonic nanostructure includes a plasmonic nanostructure core; a polymeric hydrogel attached to the plasmonic nanostructure core, the polymeric hydrogel having a thickness ranging from about 10 nm to about 200 nm; and a plurality of primers attached to side chains or arms of the polymeric hydrogel, wherein at least some of the plurality of primers are attached to the polymeric hydrogel at different distances from the plasmonic nanostructure core.

An example of a functionalized plasmonic nanostructure includes a plasmonic nanostructure core; a polymeric hydrogel attached to the plasmonic nanostructure core, the polymeric hydrogel having a thickness ranging from about 10 nm to about 200 nm; and a plurality of primers attached to side chains or arms of the polymeric hydrogel, wherein at least some of the plurality of primers are attached to the polymeric hydrogel at different distances from the plasmonic nanostructure core.

A system in an embodiment can comprise an optical assembly, an surface-plasmon-resonance (SPR) light source, and an SPR camera. The optical assembly can comprise a hemispherical prism comprising a top surface configured to support a SPR sensor; and a high numerical aperture (NA) lens located distal from the top surface of the hemispherical prism. The SPR light source can be configured to emit a light beam for SPR imaging. The SPR camera can be configured to capture an SPR image. The SPR sensor further can comprise a surface configured to contact a sample. The high NA lens can be configured to refract the light beam toward the hemispherical prism. The hemispherical prism can be configured to collimate the light beam, as refracted by the high NA lens, toward the SPR sensor. The high NA lens further can be configured to receive and refract the light beam toward the SPR camera, after the light beam is reflected by the surface of the SPR sensor. Other embodiments are disclosed.

A method for measuring molecular interactions on a plurality of regions of interest (ROIs) of a sensor surface of a biosensor device. The method can include receiving respective biosensor response data for each ROI of the plurality of ROIs. The method further can include determining a sample group and a reference group for the plurality of ROIs. The sample group can include sample group ROIs of the plurality of ROIs, and the reference group can include reference group ROIs of the plurality of ROIs. The method also can include generating one or more sample data distributions based on one or more respective sample group binding parameters for each of the sample group ROIs derived from the respective biosensor response data for the each of the sample group ROIs. The method further can include generating one or more reference data distributions based on one or more respective reference group binding parameters for each of the reference group ROIs derived from the respective biosensor response data for the each of the reference group ROIs. Other embodiments are disclosed.