Provided herein is a method of isolating cellular components from at least one region of interest in a planar tissue section.

Provided herein is a method of isolating cellular components from at least one region of interest in a planar tissue section.

A Surface-Enhanced Raman Spectroscopy (SERS) device to perform accurate label-free long-read DNA sequencing. A Raman sensor has a hot spot defined by plasmonic nanostructures and excited by at least one laser. An immobilized DNA polymerase can be used to pull a DNA template strand to be sequenced through the hot spot.

A Surface-Enhanced Raman Spectroscopy (SERS) device to perform accurate label-free long-read DNA sequencing. A Raman sensor has a hot spot defined by plasmonic nanostructures and excited by at least one laser. An immobilized DNA polymerase can be used to pull a DNA template strand to be sequenced through the hot spot.

A methodology for assays and diagnostics utilizes a nanoporous or corrugated metal-containing surface, fiber or particle which enhances or suppresses the optical detectability of a label. The resulting optical, electromagnetic, or imaging signal signals the presence of a pathogen or analyte of interest. Preferred embodiments pertain to label-free, in situ monitoring of individual DNA hybridization in microfluidics using molecular sentinel probes immobilized on nanoporous gold disks. By immobilizing molecular sentinel probes on nanoporous gold disks, single-molecule sensitivity is demonstrated via surface-enhanced Raman scattering which provides robust signals. The described methodology is generally applicable to most amplification independent assays and molecular diagnostics.

A methodology for assays and diagnostics utilizes a nanoporous or corrugated metal-containing surface, fiber or particle which enhances or suppresses the optical detectability of a label. The resulting optical, electromagnetic, or imaging signal signals the presence of a pathogen or analyte of interest. Preferred embodiments pertain to label-free, in situ monitoring of individual DNA hybridization in microfluidics using molecular sentinel probes immobilized on nanoporous gold disks. By immobilizing molecular sentinel probes on nanoporous gold disks, single-molecule sensitivity is demonstrated via surface-enhanced Raman scattering which provides robust signals. The described methodology is generally applicable to most amplification independent assays and molecular diagnostics.

Gold nanorattle probes are provided that are highly tunable, physiologically stable, and ultra-bright Raman probes for in vitro and in vivo surface-enhanced Raman scattering (SERS) applications. The nanorattles contain an essentially uniform gap between core and shell that is tunable and can range from 2 nm to 10 nm in width. This provides numerous advantages including allowing for increased loading with a variety of dye molecules that exhibit SERS in various spectral regions, including the tissue optical window for in vivo studies. In addition, the nanorattle probes provide an internal label when used in diagnostic methods to detect nucleic acids, proteins and other biotargets. The nanorattles have an essentially spherical gold metal nanoparticle core, a porous material of silver metal of an essentially uniform width surrounding the nanoparticle core that is loaded with one or more SERS reporter molecules, and an outer gold metal shell encapsulating the porous material.

Disclosed herein is a multiplexed design with three-dimensional plasmonic nanofocusing and confinement of light, demonstration of reproducible and robust single-molecule optical fingerprints using two complementary vibrational spectroscopy techniques (infrared and Raman spectroscopy), identification of respective vibrational modes which uniquely fingerprint the biomolecular species, and facile differentiation of respective fingerprints in DNA mixtures, as well as epigenetic modifications. While the nanometer scale mode volumes still prevent single letter identification of DNA sequence, we show an alternative method for identifying A, T, G, C DNA nucleotides in k-mers using sequences of these blocks as a unique and high-throughput alternative to single letter sequences (similar to binary and hexadecimal systems). Furthermore, additivity shown in single-molecule DNA mixtures and robust optical signatures can also be used in a raster-type step scan to identify single letter sequences. These results can pave the way for the development of a novel, high-throughput block optical sequencing (BOS) method.

Disclosed herein is a multiplexed design with three-dimensional plasmonic nanofocusing and confinement of light, demonstration of reproducible and robust single-molecule optical fingerprints using two complementary vibrational spectroscopy techniques (infrared and Raman spectroscopy), identification of respective vibrational modes which uniquely fingerprint the biomolecular species, and facile differentiation of respective fingerprints in DNA mixtures, as well as epigenetic modifications. While the nanometer scale mode volumes still prevent single letter identification of DNA sequence, we show an alternative method for identifying A, T, G, C DNA nucleotides in k-mers using sequences of these blocks as a unique and high-throughput alternative to single letter sequences (similar to binary and hexadecimal systems). Furthermore, additivity shown in single-molecule DNA mixtures and robust optical signatures can also be used in a raster-type step scan to identify single letter sequences. These results can pave the way for the development of a novel, high-throughput block optical sequencing (BOS) method.

Plasmonics-active nanoprobes are provided for detection of target biomolecules including nucleic acids, proteins, and small molecules. The nucleic acids that can be detected include RNA, DNA, mRNA, microRNA, and small nucleotide polymorphisms (SNPs). The nanoproprobes can be used in vito in sensitive detection methods for diagnosis of diseases and disorders including cancer. Multiplexing can be performed using the nanoprobes such that multiple targets can be detected simultaneously in a single sample. The methods of use of the nanoprobes include detection by a visible color change. The nanoprobes can be used in vivo for treatment of undesireable cells in a subject.