Second harmonic imaging nanoprobes and techniques for use thereof

• US 9,897,587 B2
• Filed: 10/24/2016
• Issued: 02/20/2018
• Est. Priority Date: 11/21/2006
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Pantazis, Periklis, Pu, Ye, Psaltis, Demetri, Hong, John H., Fraser, Scott E.
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US15/332,989
• Publication Number:
  US 20170082596A1
• Time in Patent Office: 484 Days
• Field of Search: None
• CPC Class Codes: A61B 5/0075 by spectroscopy, i.e. measuring spectra, e.g. Raman spectroscopy, infrared absorption spectroscopy A61B5/0071 takes precedence
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 20/00 Nanooptics, e.g. quantum optics or photonic crystals
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 21/636 using an arrangement of pump beam and probe beam; using the measurement of optical non-linear properties; non-linear optics per se G02F1/35
• CPC Class Codes: G01N 33/483 Physical analysis of biological material
• CPC Class Codes: G02F 1/37 for second-harmonic generation G02F1/3532 takes precedence
• CPC Class Codes: G06V 20/693 Acquisition

Abstract

Second harmonic nanoprobes for imaging biological samples and a method of using such probes to monitor the dynamics of biological process using a field resonance enhanced second harmonic (FRESH) technique are provided. The second harmonic generating (SHG) nanoprobes are comprised of various kinds of nanocrystals that do not possess an inversion symmetry and therefore are capable of generating second harmonic signals that can then be detected by conventional two-photon microscopy for in vivo imaging of biological processes and structures such as cell signaling, neuroimaging, protein conformation probing, DNA conformation probing, gene transcription, virus infection and replication in cells, protein dynamics, tumor imaging and cancer therapy evaluation and diagnosis as well as quantification in optical imaging.

Claims

1. A method of probing structures and biological processes comprising:

interspersing a probe nanostructure with a target of interest, such that the probe nanostructure interacts with the target of interest, and wherein the probe nanostructure formed of a crystal material having a unit cell with no inversion symmetry such that it generates a second harmonic emission when radiated by an excitation source;

radiating the probe nanostructure with an external excitation source such that the crystal material of the probe nanostructure generates a second harmonic emission;

detecting the second harmonic emission from the nanoprobe; and

wherein the at least one probe nanostructure generates the second harmonic emission regardless of the attachment and orientation of the probe nanostructure to the target.

2. The method of claim 1, wherein the probe nanostructure is a nanocrystal selected from the group consisting of organic, inorganic and combinations thereof.

3. The method of claim 1, wherein the target of interest is selected from the group consisting of an antigen, a peptide sequence, a nucleic acid sequence, RNA and DNA.

4. The method of claim 1, wherein the target of interest is indicative of the presence of one of either a specific medical condition or a specific pathogenesis.

5. The method of claim 1, wherein the probe nanostructure is one of either directly attached to or introduced into the target of interest.

6. The method of claim 1, wherein the probe nanostructure is conjugated to a probe molecule that is sensitive to the presence of the target of interest.

7. The method of claim 1, wherein the probe nanostructure is attached to a therapeutic agent.

8. The method of claim 1, wherein the step of radiating is conducted while the probe nanostructure is in vivo.

9. The method of claim 8, wherein the emissions from the probe nanostructure are utilized to determine the biodistribution of the target of interest.

10. The method of claim 1, wherein the target of interest is collected in one of either a liquid or solid sample.

11. The method of claim 1, further comprising interspersing an exciter nanostructure to the target of interest, said exciter nanostructure designed to produce an enhanced electrical field of a specified frequency when exposed to an excitation source, wherein the probe nanostructure generates a second harmonic resonance emission when brought within range of the enhanced electrical field of said exciter nanostructure.

12. The method of claim 1, wherein the excitation source is selected from the group consisting of continuous wave, modulated and pulsed lasers.

13. A method of field resonance enhanced second harmonic detection comprising:

attaching an exciter nanostructure to a target of interest, said exciter nanostructure designed to produce an enhanced electrical field of a specified frequency when exposed to an excitation source;

attaching a probe nanostructure to the target of interest, said probe nanostructure formed of a crystal material having a unit cell with no inversion symmetry such that it generates a second harmonic resonance emission when brought within range of the enhanced electrical field of said exciter nanostructure;

radiating the structure of interest with an external excitation source such that the crystal material of the probe nanostructure generates a second harmonic emission;

detecting the second harmonic emission from the nanoprobe; and

wherein the at least one probe nanostructure generates the second harmonic emission regardless of the attachment and orientation of the probe nanostructure to the target.

14. The method of claim 13, wherein the exciter nanostructure is a metal nanostructure selected from the group consisting of nanorods, nanospheres or nanoshells.

15. The method of claim 13, wherein the probe nanostructure and the emitter nanostructure are both attached to a single target of interest.

16. The method of claim 13, wherein the probe nanostructure is a nanocrystal selected from the group consisting of organic, inorganic and combinations thereof.

17. The method of claim 16, wherein the nanocrystal is selected from the group consisting of BaTiO₃, SiC, ZnO, ZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, GaAs, GaSb, GaP, GaN, InSb, LiNbO₃, KNbO₃, KTiOPO₄, Fe(IO₃)₃, Au, Ag, N-(4-nitrophenyl)-(L)-prolinol, urea, 4-Nitroaniline, 2-Methyl-4-nitroaniline, 3-Methyl-4-methoxy-4′

-nitrostilbene, β

-BaB₂O₄, LiB₃O₅, LiNbO₃, KTiOPO₄, AgGaS₂, AgGaSe₂, ZnGeP₂, GaSe, KH₂PO₄, NH₄H₂PO₄, KD₂PO₄, CsLiB₆O₁₀, KTiOAsO₄, KNbO₃, LiTaO₃, RbTiOAsO₄, BaTiO₃, MgBaF₄, GaAs, BiB₃O₆, K₂Al₂B₂O₇, KBe₂BO₃F₂, BaAlBO₃F₂, La₂CaB₁₀O₁₉, GdCa₄O(BO₃)₃, YCa₄O(BO₃)₃, Li₂B₄O₇, LiRbB₄O₇, CdHg(SCN)₄, RbTiOPO₄, LiInS₂, LiInSe₂, KB₅O₈.4H₂O, CsB₃O₅, C₄H₇D₁₂N₄PO₇, a-HIO₃, LiCOOH.H₂O, CsH₂AsO₄, CsD₂AsO₄, RbH₂PO₄, CsTiOAsO₄, Ba₂NaNb₅O₁₅, K₃Li₂Nb₅O₁₅, CO(NH₂)₂, LiIO₃, Ag₃AsS₃, HgGa₂S₄, CdGeAs₂, Ti₃AsSe₃.

18. The method of claim 13, wherein the excitation source is selected from the group consisting of continuous wave, modulated and pulsed lasers.