IP Library › Granted Patent US 12,196,675
Granted Patent B2
US 12,196,675 · App. 17/365,818 · Granted Jan 14, 2025

Sensitive and robust biosensing using plasmonic enhancement of fluorescence by rapid thermal annealed silver nanostructures

Inventors: Shuangming Li (Santa Clara, CA); Venkat R. Bhethanabotla (Tampa, FL)
Assignee: UNIVERSITY OF SOUTH FLORIDA
G01N21/554G01N21/6408G01N21/648G01N33/54346G01N33/54373B82Y15/00G01N33/553
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Quick Facts
Patent No.
US 12,196,675
App. No.
17/365,818
Granted
Jan 14, 2025
Kind
B2
Abstract

Chips and metallic nanostructure biosensors comprising a dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes thereon are disclosed herein. Also disclosed are methods of making and using the same.

Claims (32)

1. A chip comprising a dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes thereon, and further comprising a thin-film stabilizing overlayer deposited on the dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes, wherein the discontinuous and disordered metallic nanostructures of flat island shapes cover between 20-40% of the dielectric substrate, have an area density of between 5-600/μm 2 , and have an area between 50-250,000 nm 2 ;

wherein the thin-film stabilizing overlayer increases fluorescence enhancement of the chip; and

wherein the discontinuous and disordered metallic nanostructures of flat island shapes are prepared by rapid thermal annealing.

2. The chip of claim 1 , wherein the discontinuous and disordered metallic nanostructures of flat island shapes thereon are characterized in shape as hemi-spherical nanostructures, ellipsoidal nanostrucutures, peanut-shaped nanostructure, worm-like nanostructure, lace-like nanostructures, or any combination thereof.

3. A chip comprising a dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes thereon, and further comprising a thin-film stabilizing overlayer deposited on the dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes, wherein the discontinuous and disordered metallic nanostructures of flat island shapes cover between 20-40% of the dielectric substrate, have an area density of between 5-600/μm 2 ; have an average height between 10-60 nm, and have an area between 50-250,000 nm 2 ;

wherein the thin-film stabilizing overlayer increases fluorescence enhancement of the chip; and

wherein the discontinuous and disordered metallic nanostructures of flat island shapes are prepared by rapid thermal annealing.

4. The chip of claim 1 , wherein the discontinuous and disordered metallic nanostructures of flat island shapes are formed of silver.

5. The chip of claim 1 , wherein the thin-film stabilizing overlayer is between 2-15 nm thick.

6. A chip comprising a dielectric substrate and discontinuous and disordered metallic nanostructures of flat island shapes thereon and a thin-film stabilizing overlayer deposited on the dielectric substrate and discontinuous and the disordered metallic nanostructures of flat island shapes, wherein the stabilizing overlayer is a metal oxide or a polymer;

wherein the thin-film stabilizing overlayer increases fluorescence enhancement of the chip; and

wherein the discontinuous and disordered metallic nanostructures of flat island shapes are prepared by rapid thermal annealing.

7. The chip of claim 1 , wherein the dielectric substrate is a glass substrate or a piezoelectric substrate.

8. The chip of claim 6 , wherein the chip has an enhancement factor between 1.1-20.0 as compared to the dielectric substrate without the discontinuous and disordered metallic nanostructures of flat island shapes thereon.

9. A metallic nanostructure biosensor comprising the chip according to claim 1 and further comprising a target binding protein immobilized onto chip.

10. The metallic nanostructure biosensor of claim 9 further comprising a thin-film stabilizing overlayer, wherein the target binding protein immobilized onto the thin-film stabilizing overlayer.

11. The biosensor of claim 9 , wherein the target binding protein is a fragment crystallisable (Fc) binding protein or a lectin.

12. The biosensor of claim 9 further comprising a target probe.

13. The biosensor of claim 12 , wherein the target probe is a fluorescently labeled anti-antibody.

14. A method of fabricating a chip according to claim 1 , the method comprising: depositing a thin metallic film on a dielectric substrate; and applying rapid thermal annealing to the metallic film to produce the discontinuous and disordered metallic nanostructures of flat island shapes.

15. The method of claim 14 , wherein the metallic film is less than 25 nm thick.

16. The method of claim 14 , wherein the rapid thermal annealing occurs at a temperature lower than a melting point of the metallic film.

17. The method of claim 14 , wherein the rapid thermal annealing comprises heating the thin metallic film and the dielectric substrate up to an annealing temperate at an annealing rate, holding the thin metallic film and the dielectric substrate at the annealing temperature of an annealing time, and cooling the thin metallic film and the dielectric substrate to room temperature at a cooling rate.

18. The method of claim 14 , wherein the metallic film is formed of silver.

19. The method of claim 14 , further comprising coating the discontinuous and disordered metallic nanostructures of flat island shape with a thin stabilizing overlayer film, wherein shapes of the metallic nanostructures are retained after coating with the stabilizing overlayer.

20. The method of claim 19 , wherein the stabilizing overlayer is between 2-15 nm thick.

21. A method for forming a metallic nanostructure biosensor, the method comprising immobilizing a target binding protein onto the chip according to claim 1 .

22. The method of claim 21 , wherein the chip further comprises a thin-film stabilizing overlayer deposited on the dielectric substrate and discontinuous and the disordered metallic nanostructures of flat island shapes and wherein the target binding protein is immobilized onto the stabilizing overlayer.

23. A method for detecting of a biomolecular target in a sample, the method comprising contacting the metallic nanostructure biosensor according to claim 9 with the sample and a target probe, irradiating the metallic nanoscruture biosensor, and detecting a signal, wherein the biomolecular target has binding affinity with the immobilized binding protein and the target probe.

24. The method of claim 23 , wherein the biomolecular target is an antibody or a chitin.

25. The chip of claim 1 , wherein the discontinuous and disordered metallic nanostructures of flat island shapes comprise nanoparticles; and

wherein about 39% of the nanoparticles from the nanostructures having a 5 nm film thickness have areas in the range of 250-450 nm 2 , or about 47% of the nanoparticles from the nanostructures having a 15 nm film thickness have areas in the range of 0.025-0.045 μm 2 .

Assignments (2)
CONFIRMATORY LICENSE Recorded Jun 6, 2025
From: UNIVERSITY OF SOUTH FLORIDA
To: NATIONAL SCIENCE FOUNDATION
Reel/Frame 071496/0581 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 1, 2023
From: LI, SHUANGMING; BHETHANABOTLA, VENKAT R.
To: UNIVERSITY OF SOUTH FLORIDA
Reel/Frame 064776/0727 →
Continuity (2)
Provisional Application 63047055 · Jul 1, 2020
Related Publication 20220003674A1 · Jan 6, 2022
References Cited (53)
US 9164034B2 · Rubinstein · 2015 [cited by examiner]
Bauch, M., et al. (2014). Plasmon-Enhanced Fluorescence Biosensors: a Review. Plasmonics (Norwell, Mass.), 9, 781-799. doi: 10.1007/s11468-013-9660-5 (Year: 2014). [cited by examiner]
Jeong, Sunho et al., “Low-temperature solution-processed metal oxide thin film transistors”. “J. Mater. Chem. vol. 22, issue 4, p. 1243-1250. (2012). doi =10.1039/C1JM14452A”, (Year: 2012). [cited by examiner]
Accardo, Angelo et al. (2014) “Metal Structures as Advanced Materials in Nanotechnology”, Handbook of Nanomaterials Properties Apr. 2014. DOI:10.1007/978-3-642-31107-9_42 (Year: 2014). [cited by examiner]
Haoyan Wei et al., “From silver nanoparticles to thin films: Evolution of microstructure and electrical conduction on glass substrates” , Journal of Physics and Chemistry of Solids, vol. 70, Issue 2, (2009) https://doi.… [cited by examiner]
Agnihotri et al., “Size-controlled silver nanoparticles synthesized over the range 5-100 nm using the same protocol and their antibacterial efficacy.” Rsc Advances 4, No. 8 (2014): 3974-3983. [cited by applicant]
Alonzo-Medina et al., “Understanding the thermal annealing process on metallic thin films.” In IOP Conference Series: Materials Science and Engineering, vol. 45, No. 1, p. 012013. IOP Publishing, 2013. [cited by applicant]
Anker et al., “Biosensing with plasmonic nanosensors.” Nanoscience and Technology: A Collection of Reviews from Nature Journals (2010): 308-319. [cited by applicant]
Aslan et al., “Metal-enhanced fluorescence solution-based sensing platform.” Journal of fluorescence 14, No. 6 (2004): 677-679. [cited by applicant]
Aslan et al., “Metal-enhanced fluorescence: an emerging tool in biotechnology.” Current opinion in biotechnology 16, No. 1 (2005): 55-62. [cited by applicant]
Chen et al., “Atomic force bio-analytics of polymerization and aggregation of phycoerythrin-conjugated immunoglobulin G molecules.” Molecular immunology 41, No. 12 (2004): 1247-1252. [cited by applicant]
Chowdhury et al., “Effect of Ag—Cu alloy nanoparticle composition on luminescence enhancement/quenching.” The Journal of Physical Chemistry C 113, No. 30 (2009): 13016-13022. [cited by applicant]
Chowdhury et al., “Silver-copper alloy nanoparticles for metal enhanced luminescence.” Applied Physics Letters 95, No. 13 (2009): 131115. [cited by applicant]
Droz et al., “Influence of surface and protein modification on immunoglobulin G adsorption observed by scanning force microscopy.” Biophysical journal 67, No. 3 (1994): 1316-1323. [cited by applicant]
Dulkeith et al., “Fluorescence quenching of dye molecules near gold nanoparticles: radiative and nonradiative effects.” Physical review letters 89, No. 20 (2002): 203002. [cited by applicant]
Dulkeith et al., “Gold nanoparticles quench fluorescence by phase induced radiative rate suppression.” Nano letters 5, No. 4 (2005): 585-589. [cited by applicant]
Erickson, “Size and shape of protein molecules at the nanometer level determined by sedimentation, gel filtration, and electron microscopy.” Biological procedures online 11, No. 1 (2009): 32-51. [cited by applicant]
Geddes et al., “Metal-enhanced fluorescence.” Journal of fluorescence 12, No. 2 (2002): 121-129. [cited by applicant]
Geddes et al., “Metal-enhanced fluorescence (MEF) due to silver colloids on a planar surface: Potential applications of indocyanine green to in vivo imaging.” The Journal of Physical Chemistry A 107, No. 18 (2003): 3443… [cited by applicant]
Herminghaus et al., “Spinodal dewetting in liquid crystal and liquid metal films.” Science 282, No. 5390 (1998): 916-919. [cited by applicant]
Howarter et al., “Optimization of silica silanization by 3-aminopropyltriethoxysilane.” Langmuir 22, No. 26 (2006): 11142-11147. [cited by applicant]
Hsu et al., “Transparent displays enabled by resonant nanoparticle scattering.” Nature communications 5, No. 1 (2014): 1-6. [cited by applicant]
Hu et al., “Nanoscale pattern formation in Pt thin films due to ion-beam-induced dewetting.” Applied Physics Letters 76, No. 22 (2000): 3215-3217. [cited by applicant]
Kim et al., “Formation, structure, and reactivity of amino-terminated organic films on silicon substrates.” Journal of colloid and interface science 329, No. 1 (2009): 114-119. [cited by applicant]
Konkola, “Design and analysis of a scanning beam interference lithography system for patterning gratings with nanometer-level distortions.” PhD diss., Massachusetts Institute of Technology, 2003. [cited by applicant]
Lakowicz et al., “Radiative decay engineering: 2. Effects of silver island films on fluorescence intensity, lifetimes, and resonance energy transfer.” Analytical biochemistry 301, No. 2 (2002): 261-277. [cited by applicant]
Li et al., “Electrochemical growth of two-dimensional gold nanostructures on a thin polypyrrole film modified ITO electrode.” The Journal of Physical Chemistry B 109, No. 50 (2005): 23787-23793. [cited by applicant]
Liu et al., “Metal-enhanced immunofluorescence assays for detection of carcinoembryonic antigen.” In 2017 IEEE Sensors, pp. 1-3. IEEE, 2017. [cited by applicant]
Liu et al., “Integrating metal-enhanced fluorescence and surface acoustic waves for sensitive and rapid quantification of cancer biomarkers from real matrices.” ACS sensors 3, No. 1 (2018): 222-229. [cited by applicant]
Liz-Marzán, “Nanometals: formation and color.” Colloidal Synthesis of Plasmonic Nanometals (2004): 26-31. [cited by applicant]
Pompa et al., “Metal-enhanced fluorescence of colloidal nanocrystals with nanoscale control.” Nature nanotechnology 1, No. 2 (2006): 126-130. [cited by applicant]
Resnick et al., “Step & flash imprint lithography.” Materials Today 8, No. 2 (2005): 34-42. [cited by applicant]
Ruffino et al., “Controlled dewetting as fabrication and patterning strategy for metal nanostructures.” physica status solidi (a) 212, No. 8 (2015): 1662-1684. [cited by applicant]
Seo, Jung-Hun, Jung Ho Park, Seong-Il Kim, Bang Ju Park, Zhenqiang Ma, Jinnil Choi, and Byeong-Kwon Ju. “Nanopatterning by laser interference lithography: applications to optical devices.” Journal of nanoscience and nan… [cited by applicant]
Silverton et al., “Three-dimensional structure of an intact human immunoglobulin.” Proceedings of the National Academy of Sciences 74, No. 11 (1977): 5140-5144. [cited by applicant]
Srolovitz et al., “The thermodynamics and kinetics of film agglomeration.” Jom 47, No. 3 (1995): 31-36. [cited by applicant]
Thompson, “Solid-state dewetting of thin films.” Annual Review of Materials Research 42 (2012): 399-434. [cited by applicant]
Toma et al., “Fabrication of broadband antireflective plasmonic gold nanocone arrays on flexible polymer films.” Nano letters 13, No. 12 (2013): 6164-6169. [cited by applicant]
Vandenberg et al., “Structure of 3-aminopropyl triethoxy silane on silicon oxide.” Journal of colloid and interface science 147, No. 1 (1991): 103-118. [cited by applicant]
Vo-Dinh. “Surface-enhanced Raman spectroscopy using metallic nanostructures.” TrAC Trends in Analytical Chemistry 17, No. 8-9 (1998): 557-582. [cited by applicant]
Wang et al., “Preparation and characterization of poly (N-isopropylacrylamide) films on a modified glass surface via surface initiated redox polymerization.” Materials Letters 59, No. 14-15 (2005): 1736-1740. [cited by applicant]
Yadavali et al., “Pulsed laser dewetting of Au films: Experiments and modeling of nanoscale behavior.” Journal of Materials Research 28, No. 13 (2013): 1715-1723. [cited by applicant]
Yue et al., “Electron-beam lithography of gold nanostructures for surface-enhanced Raman scattering.” Journal of Micromechanics and Microengineering 22, No. 12 (2012): 125007. [cited by applicant]
Zabila et al., “Direct laser interference patterning: theory and application.” Acta Physica Polonica-Series A General Physics 115, No. 2 (2009): 591. [cited by applicant]
Zhang et al., “Superlenses to overcome the diffraction limit.” Nature materials 7, No. 6 (2008): 435-441. [cited by applicant]
Zhang et al., “Wavelength dependence of metal-enhanced fluorescence.” The Journal of Physical Chemistry C 113, No. 28 (2009): 12095-12100. [cited by applicant]
Zhang et al., “Photomediated synthesis of silver triangular bipyramids and prisms: the effect of pH and BSPP.” Journal of the American Chemical Society 132, No. 35 (2010): 12502-12510. [cited by applicant]
Araújo, A., et al., “Influence of the Substrate on the Mophology of Self-Assembled Silver Nanoparticles by Rapid Thermal Annealing”, The Journal of Physical Chemistry C 2016 120 (32), 18235-18242 DOI: 10.1021/acs.jpcc.6… [cited by applicant]
Lin, H., et al. (2017) A Large-Area Nanoplasmonic Sensor Fabricated by Rapid Thermal Annealing Treatment for Label-Free and Multi-Point Immunoglobulin Sensing. Nanomaterials, 7(5), 100. doi: 10.3390/nano7050100. pp. 1-1… [cited by applicant]
Tran, N., et al. (2018). Fluorescence Enhancement Using Bimetal Surface Plasmon-Coupled Emission from 5-Carboxyfluorescein (FAM). Micromachines, 9(9), 460. doi:10.3390/mi9090460. pp. 1-6. [cited by applicant]
Bauch, M., et al. (2014). Plasmon-Enhanced Fluorescence Biosensors: a Review. Plasmonics (Norwell, Mass.), 9, 781-799. doi:10.1007/s11468-013-9660-5. [cited by applicant]
Feng, T., et al. (2019). Deposition of gold nanoparticles upon bare and indium tin oxide film coated glass based on annealing process. Journal of Experimental Nanoscience, 14(1), 13-22. doi: 10.1080/17458080.2018.152039… [cited by applicant]
Li, S., & Bhethanabotla, V. R. (2021). Sensitive biosensing using plasmonic enhancement of fluorescence by rapid thermal annealed silver nanostructures. IEEE Sensors Journal, 21(14), 15917-15925. [cited by applicant]