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WO2012035545A2 - Nanoparticules de carbone non fonctionnalisées présentant des caractéristiques de fluorescence, leur procédé de préparation et leur utilisation comme agents de bio-imagerie et de détection de solvants - Google Patents

Nanoparticules de carbone non fonctionnalisées présentant des caractéristiques de fluorescence, leur procédé de préparation et leur utilisation comme agents de bio-imagerie et de détection de solvants Download PDF

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Publication number
WO2012035545A2
WO2012035545A2 PCT/IN2011/000610 IN2011000610W WO2012035545A2 WO 2012035545 A2 WO2012035545 A2 WO 2012035545A2 IN 2011000610 W IN2011000610 W IN 2011000610W WO 2012035545 A2 WO2012035545 A2 WO 2012035545A2
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Prior art keywords
carbon nanoparticles
functionalized
fluorescence
present
cells
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WO2012035545A3 (fr
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Himadri Bihari Bohidar
Pardip Kumar
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    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
    • C09K11/65—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing carbon
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
    • C01B32/00—Carbon; Compounds thereof
    • C01B32/15—Nano-sized carbon materials
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2004/00—Particle morphology
    • C01P2004/60—Particles characterised by their size
    • C01P2004/62—Submicrometer sized, i.e. from 0.1-1 micrometer
    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
    • C01P2004/00—Particle morphology
    • C01P2004/60—Particles characterised by their size
    • C01P2004/64—Nanometer sized, i.e. from 1-100 nanometer

Definitions

  • the present invention relates to non-functionalized carbon nanoparticles having fluorescent characteristics and method of preparation thereof.
  • the present invention relates to bioimaging of cells by employing non-functionalized carbon nanoparticles having fluorescent characteristics of present invention as bioimaging agents.
  • the present invention relates to sensing a solvent, particularly organic solvent, and more particularly common organic solvent by employing non-functionalized carbon nanoparticles having fluorescent characteristics of present invention as solvent sensing agent.
  • PET Positron Electronic Telescope
  • the present invention aims to solve the existing problems of existing tools and methods of imaging cells to know their status of health and identification of its stage by providing an imaging agent, which is capable of causing imaging of the cells to know their status of health and identify their stage, by comparison studies, immediately upon mixing with the cells and a method of imaging the cells by employing the same.
  • the present invention therefore, also aims to solve the existing problems of existing tools and methods of sensing of organic solvents by providing a sensing agent, which is capable of causing sensing of organic solvents, particularly of commonly used organic solvents promptly upon mixing with the solvent, by comparison studies, and a method of sensing of organic solvents by employing the same.
  • the present invention aims to overcome problems of time consumption, non-affordability and limited applicability of the existing tools and methods of imaging the cells and sensing the solvents, by providing an agent which can cause imaging of the cells or sensing of the organic solvent without wasting time and still being very economical, and hence, being affordable, and also being universally applicable.
  • main object of the present invention is to provide an imaging agent, which can cause imaging of cells to know their status of health and identify their stage, by comparison studies, immediately upon mixing with the cells, that is, without wasting time, and still being very economical, and hence, being affordable, and also being universally applicable, and a method of preparation thereof and a method of imaging a cell by employing said imaging agent.
  • Another main object of the present invention is to provide a sensing agent, which can sense the organic solvents, by comparison studies, promptly upon mixing with the solvent, that is, without wasting time, and still being very economical, and hence, being affordable, and also being universally applicable, and a method of preparation thereof and a method of sensing a solvent by employing said sensing agent.
  • Still another main object of the present invention is to provide an agent which can cause imaging of the cells or sensing of the organic solvent without wasting time and still being very economical, and hence, being affordable, and also being universally applicable, and a method of preparation thereof.
  • Yet another main object of the present invention is to provide use of agent to cause imaging of cells or sensing of organic solvents without wasting time and still being very economical, and hence, being affordable, and also being universally applicable.
  • the present invention relates to non-functionalized carbon nanoparticles having fluorescence characteristics, which are capable of causing bioimaging of cells.
  • the present invention relates to non-functionalized carbon nanoparticles having fluorescence characteristics, which are capable of causing sensing of organic solvents, particularly of commonly used organic solvents.
  • the present invention relates to method of manufacturing the non-functionalized carbon nanoparticles having fluorescence characteristics, wherein the method comprises steps of:
  • the present invention relates to method of using non- functionalized carbon nanoparticles having fluorescence characteristics for imaging the cells of sensing the solvents.
  • the present invention relates to method of imaging cells by employing non-functionalized carbon nanoparticles having fluorescence characteristics.
  • the present invention relates to method of sensing solvents by employing non-functionalized carbon nanoparticles having fluorescence characteristics.
  • Figure 1 illustrates that non-functionalized carbon nanoparticles, prepared in accordance with one of the embodiments of present invention, when dispersed in water and viewed by fluorescence microscope, emit [or show] distinct fluorescence under ultra-violet [UV]-excitation of wavelength varying from about 330 to about 385 nm [Figure IB], blue excitation of wavelength varying from about 470 to about 495 nm [Figure 1C], and green excitation of wavelength varying from about 530 to about 550 nm [ Figure ID] confirming that the non-functionalized carbon nanoparticles of present invention are capable of being imaged and to cause imaging under different excitations due to their capability to emit different and distinct fluorescence of different and distinct colour contrast under different excitations.
  • Figure 1A illustrates image of non-functionalized carbon nanoparticles of present invention under bright field.
  • Figure 2 illustrates that when cells are viewed by Laser Scanning Confocal Microscope, they show very light fluorescence due to their auto fluorescence characteristics under ultra-violet [UV] -excitation of wavelength about 405 nm [ Figure 2B], blue excitation of wavelength of about 488 nm [Figure 2C], and green excitation of wavelength of about 543 nm [ Figure 2D] confirming that the cells without non- functionalized carbon nanoparticles of present invention are not capable of causing distinguishable imaging under different excitations due to their inability to emit or produce distinguishable, different and distinct fluorescence of distinguishable, different and distinct colour contrast under different excitations.
  • Figure 2A illustrates image of cells under bright field.
  • Figure 3 illustrates that non-functionalized carbon nanoparticles, prepared in accordance with one of the embodiments of present invention, when mixed with cells and viewed by Laser Scanning Confocal Microscope, emit [show] distinguishable and distinct fluorescence under ultra-violet [UV]-excitation of wavelength about 405 nm [ Figure 3B], blue excitation of wavelength of about 488 nm [Figure 3C], and green excitation of wavelength of about 543 nm [ Figure 3D] confirming that the non- functionalized carbon nanoparticles of present invention are capable of causing imaging of cells under different excitations due to their capability to emit different and distinct fluorescence of different and distinct colour contrast under different excitations.
  • Figure 3 A illustrates image of non-functionalized carbon nanoparticles of present invention mixed with cells under bright field.
  • the cells employed in experiments of Figure 3 are human embryonic kidney (HEK-297) cells obtained from National Center for Cell Science [NCCS], India.
  • Figure 4A illustrates that non-functionalized carbon nanoparticles, prepared in accordance with one of the embodiments of present invention, when mixed in a solvent and viewed under white light and UV light of wavelength of about 300 nm, emit [or show] distinct fluorescence of distinct colour under UV light of wavelength of about 300 nm, wherein
  • Figure 4(A)(a) illustrates fluorescence of acetic acid under white light
  • Figure 4(A)(b) illustrates fluorescence of acetic acid under UV light
  • Figure 4(A)(c) illustrates fluorescence of methanol under white light
  • Figure 4(A)(d) illustrates fluorescence of methanol under UV light
  • Figure 4(A)(e) illustrates fluorescence of ethanol under white light
  • Figure 4(A)(f) illustrates fluorescence of ethanol under UV light
  • Figure 4(A)(g) illustrates fluorescence of propanol under white light
  • Figure 4(A)(h) illustrates fluorescence of propanol under UV light
  • Figure 4(A)(i) illustrates fluorescence of butanol under white light
  • Figure 4(A)(j) illustrates fluorescence of butanol under UV light
  • Figure 4(A)(k) illustrates fluorescence of toluene under white light
  • Figure 4(A)(1) illustrates fluorescence of toluene under UV light
  • Figure 4(A)(m) illustrates fluorescence of benzene under white light
  • Figure 4(A)(n) illustrates fluorescence of benzene under UV light
  • Figure 4(A)(o) illustrates fluorescence of p-xylene under white light
  • Figure 4(A)(p) illustrates fluorescence of p-xylene under UV light
  • Figure 4(A)(q) illustrates fluorescence of pyridine under white light
  • Figure 4(A)(r) illustrates fluorescence of pyridine under UV light
  • Figure 4(A)(s) illustrates fluorescence of tetrahydrofuran (THF) under white light
  • Figure 4(A)(t) illustrates fluorescence of tetrahydrofuran (THF) under UV light
  • non-functionalized carbon nanoparticles of present invention are capable of sensing the common organic liquids under different excitations due to their capability to emit different and distinct fluorescence of different and distinct color in different solvents.
  • Figure 4B illustrates that non-functionalized carbon nanoparticles, prepared in accordance with one of the embodiments of present invention, when mixed in a solvent and viewed under white light and UV light of wavelength of about 300 nm, emit [or show] distinct fluorescence of distinct colour under UV light of wavelength of about 300 nm, wherein
  • Figure 4(B)(a) illustrates fluorescence of iso-octane under white light
  • Figure 4(B)(b) illustrates fluorescence of iso-octane under UV light
  • Figure 4(B)(c) illustrates fluorescence of ethyl acetate under white light
  • Figure 4(B)(d) illustrates fluorescence of ethyl acetate under UV light
  • Figure 4(B)(e) illustrates fluorescence of acetonitrile under white light
  • Figure 4(B)(f) illustrates fluorescence of acetonitrile under UV light
  • Figure 4(B)(g) illustrates fluorescence of diethyl ether under white light
  • Figure 4(B)(h) illustrates fluorescence of diethyl ether under UV light
  • Figure 4(B)(i) illustrates fluorescence of cyclohexane under white light
  • Figure 4(B)(j) illustrates fluorescence of cyclohexane under UV light
  • Figure 4(B)(k) illustrates fluorescence of acetone under white light
  • Figure 4(B)(1) illustrates fluorescence of acetone under UV light
  • Figure 4(B)(m) illustrates fluorescence of pentane under white light
  • Figure 4(B)(n) illustrates fluorescence of pentane under UV light
  • Figure 4(B)(o) illustrates fluorescence of hepatne under white light
  • Figure 4(B)(p) illustrates fluorescence of heptane under UV light
  • Figure 4(B)(q) illustrates fluorescence of carbon tetrachloride (CC1 4 ) under white light
  • Figure 4(B)(r) illustrates fluorescence of carbon tetrachloride (CC )under UV light
  • Figure 4(B)(s) illustrates fluorescence of dimethyl sulphoxide (DMSO) under white light
  • Figure 4(B)(t) illustrates fluorescence of dimethyl sulphoxide (DMSO) under UV light
  • non-functionalized carbon nanoparticles of present invention are capable of sensing the common organic liquids under different excitations due to their capability to emit different and distinct fluorescence of different and distinct color in different solvents.
  • Figure 5(A) illustrates fluorescence spectra obtained under excitation at 225 nm of solvents mixed with non-functionalized carbon nanoparticles prepared in accordance with one of the embodiments of present invention, wherein graphs I, II, III, IV and V of Figure 5(A)(a) illustrate fluorescence spectra of acetic acid, methanol, ethanol, propanol, butanol respectively; and graphs I, II, III, IV and V of Figure 5(A)(b) illustrate fluorescence spectra of benzene, THF, toluene, p-xylene and pyridine respectively as obtained after mixing these solvents with non-functionalized carbon nanoparticles of present invention.
  • Figure 5(B) illustrates fluorescence spectra obtained under excitation at 225 nm of solvents mixed with non-functionalized carbon nanoparticles prepared in accordance with one of the embodiments of present invention, wherein graphs I, II, III, IV and V of Figure 5(B)(a) illustrate fluorescence spectra of acetone, carbon tetrachloride (CCI 4 ), pentane, heptane and cyclohexane respectively; and graphs I, II, III, IV and V of Figure 5(B)(b) illustrate fluorescence spectra of acetonitrile, diethyl ether, DMSO, iso-octane and ethyl acetate respectively as obtained after mixing these solvents with non-functionalized carbon nanoparticles of present invention.
  • graphs I, II, III, IV and V of Figure 5(B)(a) illustrate fluorescence spectra of acetone, carbon tetrachloride (CCI 4 ), pen
  • the inventors have found that when carbon nanoparticles having non-functionalized surface produced by depositing carbon soot emanating from flame of a burner on copper surface are mixed with cells, the cells, surprisingly and unexpectedly, emit [or show] distinguishable and distinct fluorescence of different and distinct colour contrast under various excitations including ultra-violet [UV] -excitation of wavelength about 405 nm, blue excitation of wavelength of about 488 nm, and green excitation of wavelength of about 543 nm, and such fluorescence from cells has been found to be capable of causing imaging of cells. It has also been observed that such image of cells may be used to know status of health of the cells being imaged and identification of its stage by comparison method.
  • UV ultra-violet
  • the inventors have also found that when carbon nanoparticles having non- functionalized surface produced by depositing carbon soot emanating from flame of a burner on copper surface are mixed with solvents, the solvents, surprisingly and unexpectedly, emit [or show] distinguishable and distinct fluorescence of different and distinct colour contrast under various excitations including ultra-violet [UV]-light of wavelength of about 300 nm, excitation of wavelength of about 225 nm, and such fluorescence from solvents has been found to be capable of causing sensing of solvents.
  • various excitations including ultra-violet [UV]-light of wavelength of about 300 nm, excitation of wavelength of about 225 nm, and such fluorescence from solvents has been found to be capable of causing sensing of solvents.
  • the “comparison method” referred herein means that image of the cell to be imaged is compared with image of the normal cell, wherein said image is obtained after mixing the cells with non-functionalized carbon nanoparticles of present invention having fluorescence characteristics.
  • the present invention relates to non-functionalized carbon nanoparticles having fluorescence characteristics, which are capable of causing bioimaging of cells.
  • the image thus obtained from the cell after mixing it with non- functionalized carbon nanoparticles of the present invention can be compared with image of a normal cell obtained from the normal cell after mixing it with non- functionalized carbon nanoparticles of the present invention to know status of health of the cell being imaged and identify its stage.
  • the cell is selected from a group comprising animal cell, human cell, wherein the human cell for example is human embryonic kidney (HEK-297) cell.
  • human embryonic kidney (HEK-297) cell for example is human embryonic kidney (HEK-297) cell.
  • the present invention relates to non-functionalized carbon nanoparticles having fluorescence characteristics, which are capable of causing sensing of organic solvents, particularly of commonly used organic solvents.
  • the fluorescence thus obtained from the solvent after mixing it with non- functionalized carbon nanoparticles of the present invention and the fluorescence spectra obtained from mixture of solvent and non-functionalized carbon nanoparticles of the present invention can be used to identify the solvent.
  • the solvent is organic solvent, wherein the organic solvent is selected from a commonly available organic solvent.
  • the organic solvent is selected from a group comprising acetic acid, methanol, ethanol, propanol, butanol, toluene, benzene, p-xylene, pyridine, tetrahydrofuran (THF), iso- octane, ethyl acetate, acetonitrile, diethyl ether, cyclohexane, acetone, pentane, hepatne, carbon tetrachloride (CC ), and dimethyl sulphoxide (DMSO).
  • acetic acid methanol, ethanol, propanol, butanol, toluene, benzene, p-xylene, pyridine, tetrahydrofuran (THF), iso- octane, ethyl acetate, acetonitrile, diethyl ether, cyclohexane, acetone, pentane,
  • non-functionalized means the surface of carbon nanoparticles of present invention is not functionalized and is used without any preparation or modifications.
  • carbon nanoparticles of present invention emit fluorescence under excitations of different wavelength, these are also referred to as “non-functionalized fluorescent carbon nanoparticles”.
  • the present invention relates to method of preparation of non-functionalized fluorescent carbon nanoparticles of present invention, wherein the method comprises steps of:- a) depositing carbon soot emanating from flame of a burner on copper surface; b) continuing step - a) till at least a layer of carbon soot is deposited on copper surface;
  • the carbon nanoparticles produced are of about 30 nm or larger, preferably of size varying from about 30 nm to about 180 nm. It has been found that method of present invention spontaneously produces non- functionalized fluorescent carbon nanoparticles of about 30 nm size. Therefore, further step to separate out the nanoparticles is not required in accordance with advantage of method of present invention.
  • step b) it is sufficient if an amount of carbon soot is deposited which is required for the intended purpose.
  • the carbon soot is generated from oil lamp provided with cotton wick.
  • the oil in oil lamp is commercially available vegetable oil selected from a group comprising mustard oil, refined vegetable oil, coconut oil and butter oil.
  • non-functionalized fluorescent carbon nanoparticles of present invention can be prepared just in one step in minimum possible time sufficient to deposit an amount required for the intended purpose.
  • the human embryonic kidney (HEK-297) cells obtained from National Center for Cell Science [NCCS], India were mixed with non-functionalized fluorescent carbon nanoparticles of present invention and viewed by Laser Scanning Confocal Microscope under ultra-violet [UV]-excitation of wavelength about 405 nm, blue excitation of wavelength of about 488 nm, and green excitation of wavelength of about 543 nm.
  • UV ultra-violet

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  • Nanotechnology (AREA)
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  • Crystallography & Structural Chemistry (AREA)
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  • General Physics & Mathematics (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne des nanoparticules de carbone non fonctionnalisées présentant des caractéristiques de fluorescence et capables de générer une bio-imagerie de cellules et de détecter des solvants organiques. Selon un mode de mise en oeuvre, l'invention concerne un procédé de fabrication dedites nanoparticules de carbone fluorescentes non fonctionnalisées consistant à: a) déposer de la suie émanant de la flamme d'un brûleur sur la surface en cuivre jusqu'à ce qu'au moins une couche de suie se dépose sur la surface en cuivre, et b) retirer la couche de suie de la surface en cuivre afin de générer des nanoparticules de carbone non fonctionnalisées présentant des caractéristiques de fluorescence, lesquelles peuvent être directement utilisées comme agent de bio-imagerie ou agent de détection de solvants.
PCT/IN2011/000610 2010-09-14 2011-09-07 Nanoparticules de carbone non fonctionnalisées présentant des caractéristiques de fluorescence, leur procédé de préparation et leur utilisation comme agents de bio-imagerie et de détection de solvants Ceased WO2012035545A2 (fr)

Applications Claiming Priority (2)

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IN2184DE2010 2010-09-14
IN2184/DEL/2010 2010-09-14

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WO2012035545A2 true WO2012035545A2 (fr) 2012-03-22
WO2012035545A3 WO2012035545A3 (fr) 2012-05-31

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105462584A (zh) * 2015-12-03 2016-04-06 安徽师范大学 荧光碳点及其制备方法和led芯片灌装组合物

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007050984A2 (fr) * 2005-10-27 2007-05-03 Clemson University Nanoparticules de carbone fluorescentes
NZ570093A (en) * 2008-07-28 2010-11-26 Auckland Uniservices Ltd Method of making luminescent nanoparticles from carbohydrates

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105462584A (zh) * 2015-12-03 2016-04-06 安徽师范大学 荧光碳点及其制备方法和led芯片灌装组合物

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