Sized-based detection techniques for detection of bio-nanoparticles are described. Detection nanoparticles and methods of forming the detection nanoparticles that can be utilized in the techniques are described. Detection nanoparticles can include modified bacteriophage that express a linking agent for a specific binding agent. Detection nanoparticles can bind a functional bio-nanoparticle with high specificity through specific binding of one or more entities unique to the functional bio-nanoparticle of interest. A detection nanoparticle can target an entity of a bio-nanoparticle that is relevant to its function, and as such, the methods can provide improvements in detection of complete and functional bio-nanoparticles. Size-based detection regimes can include particle displacement measurement techniques based upon Brownian motion.

Sized-based detection techniques for detection of bio-nanoparticles are described. Detection nanoparticles and methods of forming the detection nanoparticles that can be utilized in the techniques are described. Detection nanoparticles can include modified bacteriophage that express a linking agent for a specific binding agent. Detection nanoparticles can bind a functional bio-nanoparticle with high specificity through specific binding of one or more entities unique to the functional bio-nanoparticle of interest. A detection nanoparticle can target an entity of a bio-nanoparticle that is relevant to its function, and as such, the methods can provide improvements in detection of complete and functional bio-nanoparticles. Size-based detection regimes can include particle displacement measurement techniques based upon Brownian motion.

Methods and apparatus for long read, label-free, optical nanopore long chain molecule sequencing. In general, the present disclosure describes a novel sequencing technology based on the integration of nanochannels to deliver single long-chain molecules with widely spaced (>wavelength), ˜1-nm aperture “tortuous” nanopores that slow translocation sufficiently to provide massively parallel, single base resolution using optical techniques. A novel, directed self-assembly nanofabrication scheme using simple colloidal nanoparticles is used to form the nanopore arrays atop nanochannels that unfold the long chain molecules. At the surface of the nanoparticle array, strongly localized electromagnetic fields in engineered plasmonic/polaritonic structures allow for single base resolution using optical techniques.

Methods and apparatus for long read, label-free, optical nanopore long chain molecule sequencing. In general, the present disclosure describes a novel sequencing technology based on the integration of nanochannels to deliver single long-chain molecules with widely spaced (>wavelength), ˜1-nm aperture “tortuous” nanopores that slow translocation sufficiently to provide massively parallel, single base resolution using optical techniques. A novel, directed self-assembly nanofabrication scheme using simple colloidal nanoparticles is used to form the nanopore arrays atop nanochannels that unfold the long chain molecules. At the surface of the nanoparticle array, strongly localized electromagnetic fields in engineered plasmonic/polaritonic structures allow for single base resolution using optical techniques.

Disclosed herein are methods, compositions and systems for the interrogation of macromolecules, more particularly for preparation of isolated single macromolecules for subsequent processing of specific regions of interest within said macromolecule based on an analysis of the molecule's physical map. The disclosure is further related to the controlled segmentation of long nucleic acid parent molecules into smaller child molecules in a targeted manner such that further processing on said children may be performed with the knowledge of their origin within the parent, in a controlled environment enabled by purposefully designed microfluidic devices. Also disclosed is binding of regional specific barcodes along the length of a long nucleic acid molecule such that upon cleavage of said molecule into child molecules, the regional origin of the children can be tracked, in a controlled environment enabled by purposefully designed microfluidic devices. Finally, the disclosure is further related to droplet devices and method to control the encapsulation of long nucleic acid molecules or specific subregions thereof into a droplet, and further tracking said droplets with their contents.

Disclosed herein are methods, compositions and systems for the interrogation of macromolecules, more particularly for preparation of isolated single macromolecules for subsequent processing of specific regions of interest within said macromolecule based on an analysis of the molecule's physical map. The disclosure is further related to the controlled segmentation of long nucleic acid parent molecules into smaller child molecules in a targeted manner such that further processing on said children may be performed with the knowledge of their origin within the parent, in a controlled environment enabled by purposefully designed microfluidic devices. Also disclosed is binding of regional specific barcodes along the length of a long nucleic acid molecule such that upon cleavage of said molecule into child molecules, the regional origin of the children can be tracked, in a controlled environment enabled by purposefully designed microfluidic devices. Finally, the disclosure is further related to droplet devices and method to control the encapsulation of long nucleic acid molecules or specific subregions thereof into a droplet, and further tracking said droplets with their contents.

The subject invention provides methods and assays for multiplexed detection of analytes using nanocrystals that are uniform in morphology, size, and composition based on their unique optical characteristics. The described methods and assays are particularly useful for detection of microbes and/or microbe-based agents in a complex environmental sample.

The subject invention provides methods and assays for multiplexed detection of analytes using nanocrystals that are uniform in morphology, size, and composition based on their unique optical characteristics. The described methods and assays are particularly useful for detection of microbes and/or microbe-based agents in a complex environmental sample.

The invention provides a method for detecting the presence of a target nucleic acid analyte, for example a pathogen or virus nucleic acid, in a sample using oligonucleotide probe-functionalised nanoparticles, where hybridisation of at least three different oligonucleotide probes to at least three different target sequences in the target analyte causes agglomeration of the nanoparticles and a visible colour change. The invention also provides a population of such oligonucleotide probe-functionalised nanoparticles and a related kit for detection of a target nucleic acid analyte.

The invention provides a method for detecting the presence of a target nucleic acid analyte, for example a pathogen or virus nucleic acid, in a sample using oligonucleotide probe-functionalised nanoparticles, where hybridisation of at least three different oligonucleotide probes to at least three different target sequences in the target analyte causes agglomeration of the nanoparticles and a visible colour change. The invention also provides a population of such oligonucleotide probe-functionalised nanoparticles and a related kit for detection of a target nucleic acid analyte.