Structures for Enhancement of Local Electric Field, Light Absorption, Light Radiation, Material Detection and Methods for Making and Using of the Same.
This disclosure provides, among other things, a nanosensor comprising a substrate and one or a plurality of pillars extending from a surface of the substrate, where the pillars comprise a metallic dot structure, a metal disc, and a metallic back plane. The nanosensor comprises a molecular adhesion layer that covers at least a part of the metallic dot structure, the metal disc, and/or the metallic back plane and a capture agent bound to the molecular adhesion layer. The nanosensor amplifies a light signal from an analyte, when the analyte is specifically bound to the capture agent.
1 . A nanodevice comprising:
(a) a substrate; and
(b) one or a plurality of pillars extending from a surface of the substrate, wherein at least one of said pillars comprises:
i. a metallic disc on top of the pillar;
ii. a metallic back plane at the foot of the pillar, said metallic back plane covering a substantial portion of said substrate surface near the foot of the pillar;
iii. a metallic dot structure on sidewall of the pillar; and
iv. a molecular adhesion layer that covers at least a part of said metallic dot structure, said metal disc, and/or said metallic back plane;
wherein said nanosensor amplifies a light signal that is proximal to the exterior surface of said adhesion layer.
2 . The nanodevice of claim 1 , wherein the exterior surface of said molecular adhesion layer comprises a capture agent-reactive group, selected from an amine-reactive group, a thiol-reactive group, a hydroxyl-reactive group, an imidazolyl-reactive group and a guanidinyl-reactive group.
3 . The nanodevice of claim 2 , wherein said capture agent-reactive group is a N-hydroxysuccinimidyl ester, sulfo-N-hydroxysuccinimidyl ester, a halo-substituted phenol ester, pentafluorophenol ester, a nitro-substituted phenol ester, an anhydride, isocyanate, isothiocyanate, an imidoester, maleimide, iodoacetyl, hydrazide, an aldehyde, or an epoxide.
4 . The nanodevice of claim 1 , wherein said molecular adhesion layer is attached to said metallic dot structure, said metal disc, and/or said metallic back plane via a metal-sulfur bond.
5 . The nanodevice of claim 1 , wherein said molecular adhesion layer has a thickness of 0.5 nm to 50 nm.
6 . The nanodevice of claim 1 , wherein the molecular adhesion layer is a monolayer of alkanethiol or thio-poly(ethylene)glycol.
7 . The nanodevice of claim 1 , wherein the molecular adhesion layer is attached to said metallic dot structure, said metal disc, and/or said metallic back plane via a streptavidin/biotin interaction.
8 . The nanodevice of claim 1 , wherein the exterior surface of said molecular adhesion layer comprises a biotin moiety or streptavidin.
9 . The nanodevice of claim 1 , the exterior surface of said metallic dot structure, said metal disc, and/or said metallic back plane comprises a streptavidin group that can bind to a biotinylated capture agent.
10 . The nanodevice of claim 1 , the exterior surface of said metallic dot structure, said metal disc, and/or said metallic back plane comprises a biotin moiety that can bind to a streptavidin-linked capture agent.
11 . The nanodevice of claim 1 , wherein said nanosensor is disposed within a container.
12 . The nanodevice of claim 1 , wherein said molecular adhesion layer is a self-assembled monolayer (SAM), wherein each molecule of the SAM comprises three parts: (i) a head group that has specific affinity to the metal surfaces of the nanodevice, (ii) a terminal group that specific affinity to the capture agent, and (iii) a linker that links the head group and terminal group, wherein the length of the linker determines the average spacing between the metal surfaces and an attached capture agent can affects light amplification of the nanodevice.
13 . The nanodevice of claim 1 , wherein the metal is selected from the group consisting of gold, silver, copper, aluminum, alloys thereof, and combinations thereof.
14 . The nanodevice of claim 1 , wherein the top of said pillar has a shape selected from the group of shapes consisting of round, polygonal, pyramidal, elliptical, elongated bar shaped, or any combination thereof.
15 . The nanodevice of claim 1 , wherein the lateral dimension of said metallic disc is in the range from 5 nm to 150 nm.
16 . The nanodevice of claim 1 , wherein said metallic disc and the metallic back plane are spaced by a distance in the range of 0.1 nm to 60 nm.
17 . The nanodevice of claim 1 , wherein said at least one metallic dot structure has dimensions in the range of 1 nm to 25 nm.
18 . The nanodevice of claim 1 , wherein the distance between said metallic dot structure and said metallic disc, and the distance between said metallic dot structure and said metallic backplane is in the range of 0.5 nm to 50 nm.
19 . The nanodevice of claim 1 , wherein the spacing between the two nearest pillars of said plurality of pillars is in the range from 2 nm to 200 nm.
20 . The nanodevice of claim 1 , wherein said pillar has a sidewall surface that is columnar, sloped, or curved.
21 . The nanodevice of claim 1 , wherein the thickness of the said metallic disc and metallic back plane is between 5 nm to 60 nm.
22 . The nanodevice of claim 1 , wherein said pillar has a lateral dimension or a height less than the wavelength of said light.
23 . The nanodevice of claim 1 , wherein said metallic disc has substantially the same lateral geometry as said pillar.
24 . The nanodevice of claim 1 , wherein said pillar comprises a dielectric or semiconductor material selected from the group consisting of polymers, silicon-dioxide, silicon-nitride, hafnium oxide, aluminum oxide, silicon, gallium arsenide, and gallium nitride.
25 . The nanodevice of claim 1 , wherein the lateral dimension of said metallic disc is less than the wavelength of said light.
26 . A method of making a nanosensor, comprising:
attaching a capture agent to the molecular adhesion layer of a nanodevice of claim 1 .
27 . The method of claim 26 , wherein said capture agent is a protein or nucleic acid.
28 . The method of claim 27 , wherein said capture agent is an antibody.
29 . The method of claim 27 , wherein said capture agent is an oligonucleotide.
30 . A nanosensor, comprising:
(a) a nanodevice of claim 1 ; and
(b) a capture agent that specifically binds to an analyte, wherein said capture agent is linked to the molecular adhesion layer of said nanodevice;
wherein said nanosensor amplifies a light signal from an analyte, when said analyte is bound to said capture agent.
31 . The nanosensor of claim 30 , wherein said light signal is luminescence or fluorescence.
32 . The nanosensor of claim 30 , wherein said capture agent is a protein.
33 . The nanosensor of claim 32 , wherein said capture agent is an antibody.
34 . The nanosensor of claim 30 , wherein said capture agent is a nucleic acid.
35 . The nanosensor of claim 30 , wherein said capture agent is an oligonucleotide.
36 . The nanosensor of claim 30 , further comprising a labeled analyte that is specifically bound to said capture agent.
37 . The nanosensor of claim 36 , wherein said labeled analyte is directly or indirectly labeled with a light-emitting label.
38 . The nanosensor of claim 37 , wherein said labeled analyte is linked to said light-emitting label via a streptavidin/biotin interaction.
39 . The nanosensor of claim 30 , wherein the thickness of said molecular adhesion layer is selected to optimize the amplification of said light signal.
40 . The nanosensor of claim 30 , wherein nanosensor is in a multi-well format, wherein each well of a multi-well plate comprises a nanosensor of claim 30 , wherein the nanosensor in each of the wells comprises a different capture agent.
41 . A system comprising:
(a) a nanosensor of claim 30 ;
(b) a holder for said nanosensor;
(c) an excitation source that induces a light signal from a label; and
(d) a reader adapted to read said light signal.
42 . The system of claim 41 , wherein said excitation source is a light source.
43 . The system of claim 41 , wherein said excitation source is electrical current.
44 . The system of claim 41 , wherein said reader is a photodetector, a CCD camera, a CMOS camera, a spectrometer or an optical sensor capable of producing a two dimensional spectral map of a surface of said nanosensor.
45 . The system of claim 41 , wherein said nanosensor is in a multi-well format, and said holder and/or said reader can be moved so that reader can read a light signal from each of the wells independently.
46 . A method of detecting and/or quantifying an analyte, comprising:
(a) attaching a capture agent to the molecular adhesion layer of a nanodevice of claim 1 to produce a nanosensor;
(b) contacting a sample containing a target analyte with said nanosensor, wherein said target analyte specifically binds to said capture agent and said contacting is done under conditions suitable for said binding; and
(c) reading a light signal from any target analyte that is bound to said nanosensor;
wherein said method further comprises labeling said target analyte with a light-emitting label, either prior to or after it is bound to said capture agent.
47 . The method of claim 46 , wherein said reading applies an exciting said light emitting label using light, electricity, a chemical or combination of thereof, and measuring at least one property of said light signal selected from intensity, wavelength, and location.
48 . The method of claim 46 , wherein said analyte is labeled after it is bound to the capture agent of said nanosensor.
49 . The method of claim 46 , wherein said labeling is done by binding said target analyte to a detection agent that specifically binds to said target analyte and that is linked to a light-emitting label.
50 . The method of claim 46 , wherein said method comprise labeling said light-emitting label after it is bound to the capture agent, wherein said method further comprises removing any unbound light-emitting label from the nanosensor prior to said reading.
51 . The method of claim 49 , wherein said detection agent is a secondary antibody that comprises a light-emitting label.
52 . The method of claim 49 , wherein said detection agent is a nucleic acid that comprises a light-emitting label.
53 . The method of claim 46 , wherein said light-emitting label is a fluorescent, chemiluminescent or electroluminescent label.
54 . The method of claim 53 , wherein said light-emitting label is labeled with IRDye800CW, Alexa 790 or Dylight 800.
55 . The method of claim 46 , wherein said label emits light at a wavelength in the range of 300 nm to 1200 nm.
56 . The method of claim 46 , wherein said method comprises blocking said nanosensor prior to said contacting step (b), thereby preventing non-specific binding of said capture agents to non-target analytes.
57 . The method of claim 46 , wherein said sample is a liquid sample.
58 . The method of claim 46 , wherein said sample is a clinical sample.
59 . The method of claim 46 , wherein said sample is derived from a bodily fluid.
60 . The method of claim 46 , wherein said analyte is a protein.
61 . The method of claim 46 , wherein said capture agent and said analyte are nucleic acids.
62 . The method of claim 46 , wherein said analyte is a cancer biomarker.
63 . The method of claim 46 , wherein said analyte is a biomarker for a neurological disease.
64 . The method of claim 46 , wherein said analyte is a biomarker for cardiovascular diseases
65 . The method of claim 46 , wherein said analyte is a biomarker for organic diseases
66 . The method of claim 46 , wherein said analyte is a biomarker for an infectious or parasitic disease.
67 . A method for fabricating said nanodevice of claim 1 , comprising:
(a) patterning at least one pillar on a top surface of a substrate;
(b) depositing a metallic material layer of said top surface;
(c) allowing the metallic material deposited on the pillar tops to form a disc, the metallic material deposited on the pillar feet to form a metallic back plane, and the metallic material deposited on the sidewall to form at least one metallic dot structure;
(d) depositing a molecular adhesion layer on top of the deposited metallic material, wherein the molecular adhesion layer covers at least a part of said metallic dot structure, said metal disc, and/or said metallic back plane, and wherein the exterior surface of said molecular adhesion layer comprises a capture agent-reactive group.
68 . The method of claim 67 , further comprising:
attaching a capture agent to said molecular adhesion layer.
69 . The method of claim 67 , wherein said patterning is an embossing of a material.
70 . A method of diagnosing a disease or condition, comprising:
(a) obtaining a liquid sample from a patient suspected of having said disease or condition;
(b) contacting said sample with a nanosensor of claim 1 , wherein the capture agent of said nanosensor specifically binds to a biomarker for said disease and wherein said contacting is done under conditions suitable for specific binding of said biomarker with said capture agent;
(c) removing any biomarker that is not bound to said capture agent; and
(d) reading a light signal from biomarker that remain bound to said nanosensor, wherein a light signal indicates that said patient has said disease or condition;
wherein said method further comprises labeling said biomarker with a light-emitting label, either prior to or after it is bound to said capture agent.
71 . The method of claim 70 , wherein said patient is suspected of having cancer and said antibody binds to a cancer biomarker.
72 . The method of claim 70 , wherein said patient is suspected of having a neurological disorder and said antibody binds to a biomarker for said neurological disorder.
73 . The method of claim 70 , wherein said liquid sample comprises amniotic fluid, aqueous humour, vitreous humour, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid (CSF), cerumen (earwax), chyle, chime, endolymph, perilymph, feces, gastric acid, gastric juice, lymph, mucus (including nasal drainage and phlegm), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheum, saliva, sebum (skin oil), semen, sputum, sweat, synovial fluid, tears, vomit, urine or exhaled condensate.
74 . The method of claim 70 , wherein said sensor is employed to detect or quantify chemical compounds or biomolecules that correlate with the stage of a diseases.
75 . The method of claim 74 , wherein said disease is cancer, a cardiac disease, a pulmonary disease, a renal disease, or a mental disorder,
76 . The method of claim 70 , wherein said sensor is employed to detect or quantify a microorganism.
77 . The method of claim 76 , wherein said microorganism is a virus, fungus or bacteria from the environment or a clinical sample.
78 . The method of claim 70 , wherein said sensor is employed to detect or quantify chemical compounds or biological entities that pose hazard to food safety or national security,
79 . The method of claim 78 , wherein said chemical compounds or biological entities is toxic waste or anthrax.
80 . The method of claim 70 , wherein said sensor is employed to quantify a vital parameter in a medical or physiological monitor.
81 . The method of claim 80 , wherein said vital parameter is glucose, blood oxygen level, or total blood count.
82 . The method of claim 70 , wherein said sensor is employed to detector or quantify a specific DNA or RNA from a biosample.
83 . The method of claim 70 , wherein said sensor is employed to the sequence and compare genetic sequences in DNA in the chromosomes or mitochondria.