WO2009075768A2

WO2009075768A2 - Optical device and method for real-time chemosensitivity testing

Info

Publication number: WO2009075768A2
Application number: PCT/US2008/013371
Authority: WO
Inventors: Chad Allen Lieber; Mustafa Husen Kabeer
Original assignee: Children's Hospital Of Orange County
Priority date: 2007-12-07
Filing date: 2008-12-05
Publication date: 2009-06-18
Also published as: WO2009075768A3

Abstract

Optical systems and methods for real-time chemosensitivity testing of an in-vivo tumor are described. In one embodiment of the invention, the system comprises a probe (104) that can be inserted near a tumor. Radiation is incident on the tumor and the scattered photons from the tumor are detected by a photonic detector (107). The photonic detector (107) records a Raman spectrum which is compared to one or more Raman spectra in a database.

Description

OPTICAL DEVICE AND METHOD FOR REAL-TIME CHEMOSENSITIVITY

TESTING

CROSS-REFERENCE TO RELATED APPLICATIONS

|0001] This application claims priority to U.S. Provisional Application No. 61/012,163 filed on December 7, 2007 titled "OPTICAL DEVICE AND METHOD FOR REAL-TIME CHEMOSENSITIVITY TESTING" (Atty. Docket No. CH1HO.031PR) which is hereby expressly incorporated by reference in its entirety.

BACKGROUND Field of the Invention

[0002] This invention relates in general to optical diagnostic systems and methods. Jn particular some aspects of this invention relate to optical systems and methods for real-time chemosensitivity testing. Description of the Related Art

|0003] Cancer cells are inherently heterogeneous in bio-chemical composition and genetic content. Because of the variation resulting from the heterogeneity, a number of chemotherapeutics are available that target different components of the cell. One treatment option is to administer a selected chemotherapy to the patient and monitor the growth or recession of the tumor mass. In some cases one chemotherapeutic treatment may be more effective than the other. The process of administering a particular chemotherapeutic drug and monitoring the growth or recession of the tumor mass can be time-consuming, expensive and may delay the effective treatment of the cancer.

|0004] Of recent interest is the use of cell culture drug resistance testing (CCDRT) to determine the most effective chemotherapy for a given tumor. The word drug resistance as used herein generally refers to how well the cells survive a certain drug. The process of CCDRT begins with the harvest of cells from the tumor via biopsy or resection of the mass. The cells are then isolated and used to seed a number of cell cultures in vitro. After the cultures have reached sufficient size, which may take days or even weeks, various chemotherapies are administered to the respective cultures such that the ability of a particular chemotherapy to destroy the tumor cells can be evaluated. Once the optimum therapy is determined, it is administered to the patient. While this process improves on the efficiency of the chemotherapeutic treatment, there is an associated time and financial cost for the process. Furthermore, the behavior of the cells in culture may not be exactly the same as those inside the body, thus this technique is generally regarded with some skepticism.

|0005] Optical techniques have an increasing record of success in various biomedical applications. The advantages of these techniques are the ability to obtain morphological and/or biochemical information from a sample, and to use this information in diagnostic algorithms in rapid time frames. Raman spectroscopy is an optical technique that probes molecular vibrations of molecules. Peaks in the Raman spectrum correspond with specific chemical bonds or bond families and thus Raman spectroscopy is deemed molecular- specific. Because of its molecular specificity and relatively short acquisition times (seconds to minutes), Raman spectroscopy has been successfully utilized in a variety of diagnostic applications from pharmaceutical tablet evaluation to identification of geological samples on Mars. Raman spectroscopy also has an increasing track record in biomedical diagnostics including tissue pathology, characterizing cells and bacteria, identifying and determining biochemical content in cell nuclei, etc.

SUMMARY

|0006] Various embodiments described herein comprise systems and methods to perfoπn real-time chemosensitivity testing. In one embodiment, a Raman spectroscopy system for tumor chemosensitivity testing is disclosed. The system comprises a sample of biological material originating from the tumor: a source of electromagnetic radiation; a probe configured to be placed near or within the sample of biological material; and an analysis system, wherein the said analysis system comprises a photonic detector, a microprocessor, and diagnostic algorithms. The system is configured to identify the chemosensitivity of the tumor by comparing a Raman spectrum obtained by the system with the Raman spectra characteristic of specific drug sensitivity.

|0007] In another embodiment, a method of performing a chemosensitivity assay on a biological sample is described. The method comprises placing a probe having a sample- end, at a distance from the sample and illuminating said sample with electromagnetic radiation in certain wavelength range, through said sample-end. The method further comprises collecting photons scattered or diffusely reflected by the sample from said sample- end and acquiring a Raman spectra from the scattered or diffusely reflected photons. The acquired Raman spectrum is analyzed by comparing the acquired Raman spectra to a collection of known Raman spectra characteristic of specific drug sensitivity; and determining which of the known Raman spectra is most similar to the acquired spectrum.

BRIEF DESCRIPTION OF THE DRAWINGS

|0008] The following drawings and the associated descriptions are provided to illustrate embodiments of the present disclosure and do not limit the scope of the claims.

|0009] FlG. 1 illustrates a system for chemosensitivity testing.

[0010] FlG. 2 illustrates a probe used to perform chemosensitivity testing.

[001 1] FIG. 3 illustrated a method to perform chemosensitivity testing on a tumor in vivo

[0012] FIG. 4A illustrates a method to perform chemosensitivity testing on an excised portion of a tissue.

[0013] FlG. 4B illustrates a method to perfonn chemosensitivity testing on an excised portion of a tissue sample grown in vitro.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Although certain preferred embodiments and examples are disclosed below, inventive subject matter extends beyond the specifically disclosed embodiments to other alternative embodiments and/or uses of the invention, and to modifications and equivalents thereof. Thus, the scope of the inventions disclosed herein is not limited by any of the particular embodiments described below. For example, in any method or process disclosed herein,, the acts or operations of the method or process may be performed in any suitable sequence and are not necessarily limited to any particular disclosed sequence. For purposes of contrasting various embodiments with the prior art, certain aspects and advantages of these embodiments are described. Not necessarily all such aspects or advantages are achieved by any particular embodiment. Thus, for example, various embodiments may be carried out in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other aspects or advantages as may also be taught or suggested herein. The systems and methods discussed herein can be used anywhere, including, for example, in laboratories, hospitals, healthcare facilities, intensive care units (ICUs), or residences. Moreover, the systems and methods discussed herein can be used for invasive techniques, as well as non-invasive techniques or techniques that do not involve a body or a patient.

|0015] FIG. 1 illustrates a system for optical chemosensitivity testing. The system comprises a source of electromagnetic radiation 101 , a probe system 104 further comprising elements that can deliver electromagnetic radiation to and collect scattered photons from a sample 106, a radiation detector 107 and a micro-processor based analysis system 108. In various embodiments, the radiation detector 107 may comprise a photon detector. The system illustrated in FlG. 1 is configured to direct electromagnetic radiation in one or more wavelength ranges from the source of electromagnetic radiation 101 towards a sample 106. In some embodiments, the bandwidth of the electromagnetic radiation can be in the range from 0.01 nanometer to 5 nanometer. The scattered photons from the sample can be collected by the system illustrated in FlG. 1 and analyzed to obtain information regarding the sample. In some embodiments, the sample can be a biological material comprising one or more cells. In some embodiments, the sample can be a cross-section of bodily tissue that is harvested from a human being or animal. In some other the sample can be a mass of bodily tissue inside the body of a patient. In some embodiments, the sample can be present both in- vitro and in-vivo.

[0016] The system illustrated in FlG. 1 can be used in hospitals, urgent care centers, emergency rooms, homes, laboratories, etc. The system can be mobile and easily portable. The system can be used and operated by nurses, doctors, residents and the patients. In some embodiments, the system can be automated and designed in such a manner that it can be operated by an approximately untrained operator. In some embodiments, the system can be setup and operated in a relatively short duration.

|0017] In one embodiment, the source of electromagnetic radiation 101 can be a laser. The source 101 can comprise a light-emitting diode, a laser diode, an external cavity laser, etc. In some embodiments, the source 101 may comprise solid state, gas or semiconductor gain medium. In some embodiments, the source 101 can emit electromagnetic radiation in broadband spectral range, a continuous spectral range or in one or more discrete wavelength regions. In some embodiments, the source 101 can be monochromatic with a center wavelength between 200 and 1 100 nanometer. For example, in some embodiments, the source 101 can be centered at 633 nanometers, 785 nanometers, 830 nanometers, 900 nanometers, 980 nanometers. 930 nanometers or 1064 nanometers. The source 101 can be substantially stable and may have relatively low intensity and phase noise. For example, the bandwidth of the laser center line can be less than or equal to 5 nanometer. In some embodiments, the bandwidth of the laser center line can be less than or equal to 0.1 nanometer. In some embodiments, the source of electromagnetic radiation 101 can emit light, heat or both. In some embodiments, the source of electromagnetic radiation 101 can emit other types of radiation such as high energy particles. In some embodiments the wavelength of the electromagnetic radiation 101 can vary between 400 nanometers and 2000 nanometers.

[0018] In some embodiments, the source of electromagnetic radiation 101 can be operated in a continuous manner or in a pulsed mode. In some embodiments, the source of electromagnetic radiation 101 can be supplied electric power from an electrical power supply line. In alternate embodiments,, electrical power to the source of electromagnetic radiation 101 can be supplied by a voltage regulator. In some other embodiments, electrical power to the source of electromagnetic radiation 101 can be supplied by a battery pack. In the system illustrated in FlG. 1. the source of electromagnetic radiation can be controlled by an external controller 102. In some embodiments, the external controller 102 can switch the source of electromagnetic radiation 101 on or off. In some embodiments, the external controller 102 can be used to alternate between continuous and pulsed mode of operation. The external controller 102 can also be used to change the wavelength and/or the power of the electromagnetic radiation emitted by the source 101.

|0019] The electromagnetic radiation emitted by the source 101 can be delivered to a probe 104 by a waveguide such as an optical fiber 103. In some embodiments, micro- optical components such as micro lens that collimate and focus radiation can be used to couple radiation from the source 101 in to an optical fiber 103. In some embodiments, components such as electromagnetic radiation filters, beam splitters, mirrors, polarizers, prisms, lenses, etc. can be placed in the radiation path from the source 101 to the probe 104. [0020] The probe 104 is configured to transmit radiation from the source 101 to a sample of biological tissue 106 and collect the photons scattered or diffusely reflected by the sample 106 after interaction. The probe 104 may comprise at least one of an endoscope, a laparoscopic system and a scalpel. The probe 104 may be disposed at a distance of approximately 0 cm to approximately 30 cm from the sample 106. In some embodiments, the probe 104 may be placed in contact with the sample 106. In some embodiments, the probe 104 may be inserted into the sample 106. In some embodiments, the probe 104 may comprise components to substantially couple radiation from the source 101 to the sample 106. These components may be selected from a list comprising lens, prisms, collimators, focusers, filters, etc. In some embodiments, the components used to substantially couple radiation from the source 101 to the sample 106 may comprise miniaturized optical components such as microlens. microprisms, etc. The probe 104 may include zoom or pan features to allow movement of the measurement location independent of the probe itself. The zoom and pan settings can be controlled either mechanically or electro-mechanically or by some other technique.

[0021J Referring to FIG. 1 , the radiation that is scattered from the sample can be collected by the probe 104, and directed towards a photonic detector 107. In some embodiments, the components that collect the scattered photons can comprise high numerical aperture components that increase the photonic gathering efficiency. The photonic detector 107 can comprise an opto-electronic device such as photodiodes, charge-coupled device (CCD), photodiode arrays, complementary metal oxide semiconductor (CMOS) detectors, photomultiplier tubes, etc. In some embodiments, the photonic detector 107 may comprise a diffractive element. The diffractive element can include a reflective or transmissive grating, a prism or some other optical component that can spatially separate the different frequencies in the collected photons. In some embodiments, the spectral detector 107 may comprise a spectrometer. The information from the photonic detector 107 can be forwarded to an analysis system 108 where the information from the photonic detector 107 can be processed. In some embodiments, the analysis system 108 can comprise a microprocessor. The processing in the analysis system 108 can occur via software written for use on a computer, microcomputer or by algorithms written directly into processing chips (such as erasable programmable read only memory (EPROM)).

|0022] FlG. 2 illustrates a detailed schematic of an embodiment of the probe 104. In the embodiment shown in FlG. 2, the probe 104 comprises a beam splitter or a partially reflecting mirror 201. Radiation from the source (e.g. source 101 ) may be coupled into the probe 104 along the path 204. The incident radiation may be focused on to the sample 106 using lens assemblies 202 and 203. The incident radiation may be scattered or diffusely reflected by the sample as shown by the rays 206. In some embodiments, the lens assemblies 202 and 203 can comprise high numerical components to collect the photons that are scattered or diffusely reflected by the sample 106. In some embodiments, the collected photons 207 can be deflected along a specified path 208 by the beam splitter or the partially reflecting mirror 201. In some embodiments a separate probe may be used for collecting the scattered photons.

|0023] FlG. 3 illustrates a preferred method of use of the chemosensitivity testing system illustrated in FlG. 1. In step 303. electromagnetic radiation from the source (e.g., source 101 of FIG. 1 ) impinges on a sample (e.g., sample 106 in FIG. 1 ). In one embodiment, impinging electromagnetic radiation on the sample from the source may comprise focusing an incident beam of light on the sample by using the optical assembly (e.g.. 202 and 203 of FIG. 2) in the probe 104. The size of the focused spot may vary from 0.1 μm to 30 mm. In some embodiments, the size of the focused spot and the location of the focused spot can be controlled by the zoom and pan settings of the probe 104. In some embodiments, a spectral filter may be disposed in the path of the incident beam of light to substantially reduce scatter from the system optics. In some embodiments, the electromagnetic radiation that impinges on the sample can interact with the sample. The characteristics of the electromagnetic radiation may be altered by the interaction with the sample. For example, interaction with the sample can change the frequency spectrum or the intensity of the electromagnetic radiation. In some embodiments, upon interaction with the sample, the electromagnetic radiation can be scattered from the sample. The scattered photons after interaction with the sample can be collected by the probe 104 as shown in step 304. In some embodiments, collection optics such as high numerical aperture lenses, prisms, etc can be used to collect the scattered or diffuse reflected photons from the sample. In step 305, the collected scattered or diffusely reflected photons are detected with the photonic detector 107. The photonic detector 107 can measure and record the spectrum, intensity or other characteristics of the electromagnetic radiation after interaction with the sample. The information recorded by the photonic detector 107 can then be analyzed by an analysis system 108.

10024] In some embodiments, the photons scattered from the sample can be Raman scattered. The analysis system 108 can compare the recorded Raman spectrum from the sample to one or more known Raman spectra from a database as shown in step 306. In some embodiments, the recorded Raman spectrum may be compared to one or more Raman spectra of tissues having known chemosensitivities. In some embodiments, the database may contain the Raman spectra of different samples and the corresponding chemosensitivity. The word chemosensitivity as used herein refers generally to the growth inhibition of tumor cells after administration of an anti-cancer drug. It is understood that the phrases chemosensitivity and drug-resistance are antonyms, but that both terms are used throughout to describe the process of drug selection to maximize tumor elimination.

|0025] In some embodiments, the anti-cancer drugs may work by several different cellular interactions with the tissue. These interactions may generate unique Raman spectral features that can be evaluated in the sample. For example, a tissue s chemosensitivity to Tamoxifen can be tested by evaluating whether or not the tissue has estrogen receptors. Thus, in these embodiments, it may be advantageous to compare the recorded tissue Raman spectrum to the Raman spectrum of estrogen receptors in the database to test the chemosensitivity of the tissue to Tamoxifen. In some embodiments, the reference database (to which the recorded spectrum can be compared) may comprise information about certain features that are independent of tissue type (for example, Raman spectrum of the estrogen receptors) that could provide chemosensitivity in addition to the Raman spectra of tissues with known chemosensitivity.

|0026] The comparison can be performed by taking a ratio of the recorded Raman spectrum to one or more Raman spectra in the database. In some embodiments, comparison may be performed by calculating a cross correlation function between the recorded Raman spectrum and one or more Raman spectra in the database. Other statistical techniques such as regression analysis can also be used to compare the recorded Raman spectrum to one or more Raman spectra in the database. In some embodiments, the intensity of the Raman scattered photons at one or more wavelengths can be compared to one or more Raman spectra in the database. Based on the comparison, the Raman spectra that are most similar to the recorded Raman spectrum are selected and the corresponding chemosensitivity is noted.

(0027) In some embodiments, enhancement of the Raman scattered radiation can be employed, for example, by using surface enhanced Raman spectroscopy (SERS). In some embodiments, the surface at or near the tip of the probe (e.g., probe 104 of FlG. 1) may comprise a patterned surface. In some embodiments, the patterned surface may be metallic comprising metals such as gold, silver, etc. In some embodiments, the patterned surface may comprise a material having a crystalline structure, for example, Klarite™, silicon or some other material. In some embodiments, the patterned surface may comprise a micro-surface. In some embodiments, the patterned surface may be roughened (e.g. by an abrasive material such as sandpaper), to create indeterminate surface features on a micro scale. In some embodiments, the patterned surface may enhance the local electromagnetic and chemical fields and enhance the resulting Raman scattering cross-section in regions very near to the sample or substrate such as the cell membrane and its receptors.

|0028] In some embodiments, the enhancement can be performed with the use of nanoparticles. In some embodiments, the nanoparticles may be spherical or oblong in shape or have specific geometry as a variation thereof. For example, in some embodiments the nanoparticles may be star-shaped. In various embodiments, the nanoparticles may comprise metals such as gold or silver. In various embodiments, the size of the nanoparticles may range from approximately 1 nanometer to approximately 1 micrometer. In some embodiments, the size of the nanoparticles may lie outside this range. In some embodiments, the nanoparticles will be conjugated with specific molecules. For example, in some embodiments, the nanoparticles may be conjugated with estrogen. In some embodiments, the nanoparticles can be introduced to the cells or tissue, such that the cells or tissue uptake the particles into the interior of the cell. In some embodiments, the uptake may occur artificially via microinjection. In some embodiments, a specific target molecule may be attached to the nanoparticle. so the cells' surface receptors cndocytose it (receptor-mediated endocytosis). . In some embodiments, the uptake may occur naturally via membrane permeation or phagocytosis. These nanoparticles may enhance the Raman scattering cross-section of interior cellular elements such as the DNA, RNA, and mitochondria.

10029] In some embodiments, the above described method can be performed in- vivo, wherein the sample can be a portion of an intact tumor. FIG. 4A describes an embodiment of a method of performing chemosensitivity test using the system described in FIG. 1. In some embodiments, a portion of the intact tumor or tissue can be excised or dissected to obtain a sample as shown in step 401 of FIG. 4A. Raman spectrum of the excised portion of the tissue can be obtained in-vivo or ex-vivo as shown in step 402 of FIG. 4A. In some embodiments, the Raman spectrum of the intact tissue can be obtained in-vivo. In some embodiments, the excised portion can be removed from the body and grown in-vitro as shown in step 403 of FlG. 4B and the Raman spectra of the grown tissue can be obtained in-vitro. However, one advantage of some embodiments of the present invention is that no culture of cells or tissue is necessary. Referring to FlG. 4A, the method described above with respect to FlG. 3 is followed to obtain chemosensitivity of the sample. In some embodiments, chemosensitivity can be performed on a portion of the intact tumor in-vivo and also on an excised portion of the tumor ex-vivo. When chemosensitivity is performed both in-vivo and ex-vivo. then the Raman spectrum of the in-vivo sample can be compared to the Raman spectrum of the ex-vivo sample to correlate the in-vivo and ex-vivo samples as shown in step 407 of FlG. 4A.

|0030] In some embodiments the system and the method described above can be used to obtain Raman spectrum from in-vivo and/or ex-vivo samples after administering chemotherapy. Jn some embodiments, Raman spectra can be obtained at several locations along the sample to obtain information regarding heterogeneity of the sample

|003J | Although this invention has been disclosed in the context of certain preferred embodiments and examples, it will be understood by those skilled in the art that the present invention extends beyond the specifically disclosed embodiments to other alternative embodiments and/or uses of the invention and obvious modifications and equivalents thereof. In addition, while several variations of the invention have been shown and described in detail, other modifications, which are within the scope of this invention, will be readily apparent to those of skill in the art based upon this disclosure. It is also contemplated that various combinations or sub-combinations of the specific features and aspects of the embodiments may be made and still fall within the scope of the invention. It should be understood that various features and aspects of the disclosed embodiments can be combined with, or substituted for, one another in order to form varying modes of the disclosed invention. Thus, it is intended that the scope of the present invention herein disclosed should not be limited by the particular disclosed embodiments described above, but should be determined only by a fair reading of the claims that follow.

Claims

WHAT IS CLAIMED IS:

1. A Raman spectroscopy system for tumor chemosensitivity testing, said system comprising: a sample of biological material originating from the tumor; a source of electromagnetic radiation; a probe configured to be placed near or within the sample of biological material; and an analysis system, wherein the said analysis system comprises a photonic detector, a microprocessor, and diagnostic algorithms; said system configured to identify the chemosensitivity of the tumor by comparing a Raman spectrum obtained by the system with the Raman spectra characteristic of specific drug sensitivity.

2. The Raman spectroscopy system of Claim 1. wherein the sample of biological material comprises one or more cancer cells.

3. The Raman spectroscopy system of Claim 1. wherein the sample is selected from the group consisting of an intact tumor, a portion of the tumor or cells grown from a tumor.

4. The Raman spectroscopy system of Claim 1 , wherein the source of electromagnetic radiation is a stable laser source.

5. The Raman spectroscopy system of Claim 1. wherein the source of electromagnetic radiation is configured to emit radiation between 200 nm and 2000 nm.

6. The Raman spectroscopy system of Claim 1. wherein radiation from the source of electromagnetic radiation is delivered to the probe by an optical system.

7. The Raman spectroscopy system of Claim 6. wherein the optical system comprises optical components selected from a group consisting of lenses, filters, prisms and gratings.

8. The Raman spectroscopy system of Claim 1 , wherein the probe is configured to be placed within the sample of biological material.

9. The Raman spectroscopy system of Claim 1 , wherein the probe further comprises an optical system to deliver light from the source of electromagnetic radiation to the sample of biological material.

10. The Raman spectroscopy system of Claim 9, wherein the optical system comprises optical components selected from a group consisting of lenses, filters, prisms and gratings.

1 1. The Raman spectroscopy system of Claim 1 , wherein the photonic detector comprises a spectrometer.

12. The Raman spectroscopy system of Claim 1 , wherein a portion of the surface of the probe is patterned.

13. The Raman spectroscopy system of Claim 12, wherein the patterned surface comprises a metal.

14. The Raman spectroscopy system of Claim 12. wherein the patterned surface comprises a material having a crystalline structure.

15. The Raman spectroscopy system of Claim 12, wherein the patterned surface comprises silicon.

16. The Raman spectroscopy system of Claim 12, wherein the patterned surface comprises Klarite.

17. A method of performing a chemosensitivity assay on a biological sample, the method comprising: placing a probe having a sample-end, at a distance from the sample; illuminating said sample with radiation in one or more wavelength ranges, through said sample-end; collecting photons scattered by the sample from said sample-end; acquiring a Raman spectra from the scattered or diffusely reflected photons: analyzing the acquired Raman spectra: comparing said acquired Raman spectra to a collection of known Raman spectra characteristic of specific drug sensitivity; and determining which of the known Raman spectra is most similar to the acquired spectrum.

18. The method of Claim 17, further comprising; excising a portion of biological material said portion comprising cells; and growing said cells in-vitro to obtain said biological sample;

19. The method of Claim 17, wherein comparing the acquired Raman spectra to a collection of known Raman spectra characteristic of specific drug sensitivity comprises taking a ratio of the acquired Raman spectra to the known Raman spectra at one or more wavelengths.

20. The method of Claim 17, wherein comparing the acquired Raman spectra to a collection of known Raman spectra characteristic of specific drug sensitivity comprises calculating a cross correlation function between the acquired Raman spectra and the known Raman spectra.

21. The method of Claim 17, wherein comparing the acquired Raman spectra to a collection of known Raman spectra characteristic of specific drug sensitivity comprises a regression between the acquired Raman spectra and the known Raman spectra.

22. The method of Claim 17, wherein nanoparticles are introduced into the sample.

23. The method of Claim 22, wherein the nanoparticles are introduced by microinjection.

24. The method of Claim 22, wherein the nanoparticles are introduced by receptor-mediated endocytosis.

25. The method of Claim 22. wherein the nanoparticles are introduced by phagocytosis.

26. The method of Claim 22, wherein the nanoparticles comprise a metal.

27. The method of Claim 22, wherein the nanoparticles have a size in the range from 1 nanometer to 1 micrometer.