CN113466015B - Staining reagent for distinguishing normal cells from cancer cells, and preparation method and application thereof - Google Patents
Staining reagent for distinguishing normal cells from cancer cells, and preparation method and application thereof Download PDFInfo
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- 201000011510 cancer Diseases 0.000 title claims abstract description 35
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 33
- 239000012128 staining reagent Substances 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title abstract description 7
- MPVDXIMFBOLMNW-ISLYRVAYSA-N 7-hydroxy-8-[(E)-phenyldiazenyl]naphthalene-1,3-disulfonic acid Chemical compound OC1=CC=C2C=C(S(O)(=O)=O)C=C(S(O)(=O)=O)C2=C1\N=N\C1=CC=CC=C1 MPVDXIMFBOLMNW-ISLYRVAYSA-N 0.000 claims abstract description 31
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims description 53
- 239000011521 glass Substances 0.000 claims description 18
- 238000003384 imaging method Methods 0.000 claims description 9
- 238000010186 staining Methods 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 6
- 238000005868 electrolysis reaction Methods 0.000 claims description 6
- 238000004043 dyeing Methods 0.000 claims description 5
- 230000005284 excitation Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 239000003792 electrolyte Substances 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 2
- 230000008569 process Effects 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 2
- 238000001514 detection method Methods 0.000 abstract description 7
- 239000012530 fluid Substances 0.000 abstract 2
- 210000004027 cell Anatomy 0.000 description 106
- 210000003855 cell nucleus Anatomy 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 238000002189 fluorescence spectrum Methods 0.000 description 4
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 3
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 210000002200 mouth mucosa Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MCTQNEBFZMBRSQ-UHFFFAOYSA-N (3-amino-4-phenyldiazenylphenyl)azanium;chloride Chemical compound Cl.NC1=CC(N)=CC=C1N=NC1=CC=CC=C1 MCTQNEBFZMBRSQ-UHFFFAOYSA-N 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 206010043276 Teratoma Diseases 0.000 description 1
- 208000008383 Wilms tumor Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000009194 climbing Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000001839 endoscopy Methods 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 208000022013 kidney Wilms tumor Diseases 0.000 description 1
- 201000000289 malignant teratoma Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000008026 nephroblastoma Diseases 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000588 tumorigenic Toxicity 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
Abstract
The invention belongs to the technical field of biomedical detection, and particularly relates to a staining reagent for distinguishing normal cells from cancer cells, and a preparation method and application thereof. According to the invention, the orange G solution is used as a carbon source, and an electrolytic method is adopted to prepare the orange G carbon dot solution, wherein the orange G carbon dot solution is a staining reagent for distinguishing normal cells from cancer cells. And (3) placing the detected cell fluid treated by the staining reagent and the normal cell fluid under a fluorescence differential interference microscope, and comparing cell morphology to judge whether the detected cell is cancerous or not. The method has the advantages of simplicity and rapidness.
Description
Technical Field
The invention belongs to the technical field of biomedical detection, and particularly relates to a staining reagent for distinguishing normal cells from cancer cells, and a preparation method and application thereof.
Background
Medically, cancer refers to malignant tumors originating in epithelial tissue, the most common of which. Correspondingly, malignant tumors originating from mesenchymal tissue are collectively referred to as sarcomas. There are a few malignant tumors which are not named according to the above principles, such as nephroblastoma, malignant teratoma, etc. The term "cancer" is generally used to refer to all malignant tumors. Tumors are novel organisms formed by abnormal proliferation and differentiation caused by the fact that cells of local tissues lose normal regulation on the gene level of the organisms under the action of various tumorigenic factors. Once a new organism is formed, it does not stop growing due to the elimination of the cause, and its growth is not physiologically regulated by the normal organism, but destroys normal tissues and organs, which is particularly evident in malignant tumors.
Normally, the size and morphology of cells from the same tissue are substantially uniform, and cancer cells are generally larger than corresponding normal cells, and the size and morphology of cancer cells are also quite different from each other, and sometimes "tumor giant cells" with a very large volume are generated. The cells contain cell nuclei and cell plasma, and most cells have only one cell nucleus under normal conditions, and the cell nuclei and the cell plasma have a certain proportion; the cell nucleus of the cancer cells is enlarged in volume and inconsistent in morphology, and megakaryons, dinuclear, polynuclear or heteronuclear can appear; cancer cells grow rapidly, with obvious nuclei, whereas normal cells have no obvious nuclei.
There are many methods for detecting cancer cells, such as B-ultrasound, CT, nuclear magnetic resonance examination, and blood drawing tumor markers, puncture biopsy, endoscopy, etc. The method for judging whether the canceration phenomenon exists or not according to the form of the detected cells by using the coloring agent for distinguishing the normal cells and the detected cells of the same tissue is rarely reported in the prior art.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a staining reagent for distinguishing normal cells from cancer cells, and a preparation method and application thereof, and aims to judge whether the detected cells are cancerous or not by comparing cell morphology through placing detected cell sap treated by the staining reagent and normal cell sap under a fluorescence differential interference microscope, thereby having the advantages of simplicity and rapidness.
A staining reagent for distinguishing normal cells from cancer cells is an orange G carbon dot solution, wherein the mass concentration of orange G is 0.125%, the fluorescence excitation wavelength is 517nm, and the emission wavelength is 304nm.
The preparation method of the staining reagent for distinguishing normal cells from cancer cells comprises the following steps:
(1) PBS buffer solution is used as electrolyte to prepare an orange G solution with the concentration of 0.002 mol/L;
(2) And (3) electrolyzing the orange G solution serving as a carbon source, and when the solution gradually changes from non-fluorescence to yellow-green fluorescence, obtaining the orange G carbon dot solution.
Wherein, the PBS buffer solution pH=7.
The voltage of the electrolysis process is 10V, and the electrolysis time is 8min.
The invention also provides application of the staining reagent for distinguishing normal cells from cancer cells in cancer cell detection.
A method for detecting cancer cells, comprising the steps of:
(1) Staining of the examined cells: taking 100 mu L of detected cells, adding 1mL of PBS solution for rinsing, centrifuging at 3000rpm for 5min, discarding supernatant, replacing new PBS solution to a volume of 1mL, sucking 20-50 mu L of orange G carbon dots with mass concentration of 0.125%, adding the PBS solution, incubating for 10min, sucking, rinsing with PBS solution, and sucking to obtain detected cell liquid;
(2) Normal cell staining: adding normal cells of the same tissue into a centrifuge tube filled with PBS buffer solution, centrifuging, discarding supernatant, sucking 20-50 mu L of the orange G carbon dot solution, placing into a centrifuged cell tube, incubating for 10min, sucking, rinsing with PBS solution, discarding supernatant, and reserving the normal cell solution with suspended bottom;
(3) Dropping 100 mu L of examined cell liquid on the glass slide, then reversely buckling the stained examined cell glass slide on the glass slide, and placing the glass slide under a fluorescence microscope to observe cell imaging; simultaneously sucking 100 mu L of normal cell solution after dyeing treatment to drop on a glass slide, and placing the glass slide under a fluorescence microscope to observe cell imaging; and judging whether the detected cells are cancerous or not through the imaging comparison of the two groups of cells.
Wherein, the PBS solution pH=7.2-7.4.
Compared with the prior art, the invention has the characteristics and beneficial effects that:
the staining reagent for distinguishing normal cells from cancer cells provided by the invention is characterized in that the cell sap to be detected after staining and the normal cell sap are placed under a fluorescence differential interference microscope, and the cell morphology showing fluorescence is compared, so that whether the cell to be detected is cancerous or not can be judged.
The cancer cell detection method is simple and can be detected under a common optical microscope.
Drawings
FIG. 1 is a graph of the ultraviolet and fluorescence spectra of an exocarpium Citri Grandis G carbon dot solution of the present invention;
(a) An ultraviolet-visible spectrum; (b) is a fluorescence spectrum;
wavelength: a wavelength; absorpance: absorption rate; PL intensity: fluorescence intensity; EX: fluorescence emission wavelength; EM: fluorescence excitation wavelength; chrysoidine G CDs: orange G carbon dots;
FIG. 2 is a TEM electron micrograph of an exocarpium Citri Grandis G carbon dot solution according to the invention;
FIG. 3 is a comparison of cell imaging of stained cells and normal cells;
(a1) A cell morphology map showing the cells under test in the open field; (a2) A cell morphology map of the test cells under fluorescent conditions; (b 1) a cell morphology map of normal cells under an open field; (b2) Shows the cell morphology of normal cells under fluorescent conditions.
Detailed Description
In order to better understand the technical solution in the embodiments of the present invention and to make the above objects, features and advantages of the present invention more obvious, the following detailed description of the present invention will be given with reference to the accompanying drawings.
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and should be considered as specifically disclosed herein.
Example 1
Preparation of orange G carbon dots:
in this example, orange G (C 12 H 12 N 4 HCl) is used as a carbon source to prepare the exocarpium citri rubrum G carbon point solution by adopting an electrolysis method, wherein the electrolysis device is oneA25 mL glass bottle container and two platinum electrodes fixed on a bottle cap and 4cm long were used, and the voltage was 220V/10V.
An orange G solution with the concentration of 0.002mol/L is prepared by using PBS buffer solution (pH=7) as electrolyte, then 16ml of the orange G solution is added into an electrolytic glass bottle, an electrode is inserted and electrified for 8min, the solution gradually changes from non-fluorescent to yellow-green fluorescent solution at the beginning, which indicates carbon dot formation, and the orange G carbon dot solution is prepared.
As shown in fig. 1 and 2, wherein fig. 1 is an ultraviolet and fluorescence spectrum of the orange G carbon dot solution of the present invention, and fig. 2 is a TEM electron microscope of the orange G carbon dot solution of the present invention. Wherein FIG. 1a shows an ultraviolet-visible spectrum; FIG. 1b shows a fluorescence spectrum, and as can be seen from FIG. 1b, the fluorescence excitation wavelength of the orange G carbon dot solution is 517nm, and the emission wavelength is 304nm.
Example 2
Cell staining:
staining of cancer cells: washing cultured liver cancer cell HepG2 climbing tablet with PBS solution (pH=7.2-7.4) for 2 times, sucking 1.5ml of prepared exocarpium Citri rubrum G carbon dot solution into cells, incubating for 10min, sucking, washing with PBS solution for two times, and sucking. And (3) dripping 10 mu L of cell culture solution on the glass slide, then inversely buckling the treated cell glass slide on the glass slide, and observing cell imaging under a fluorescence microscope.
Oral epithelial cell staining: taking oral mucosa cells with a clean cotton swab, immersing the oral mucosa cells twice in a centrifuge tube filled with 1.5ml PBS buffer (pH=7.2-7.4), centrifuging at 500rpm for 6min, discarding supernatant, sucking 1.5ml of the prepared orange G carbon dot solution into a centrifuged cell tube, incubating for 10min, sucking, rinsing with PBS solution twice, discarding supernatant, and leaving cell solution in the bottom suspension part. Then, 30. Mu.L of the treated cell solution was pipetted onto a slide glass and subjected to fluorescent micromirror for cell imaging.
Example 3
Microscopic observation test:
and (3) placing the cancer cells treated by the orange G carbon point solution and the oral epithelial cell slide under a fluorescence microscope, finding clear cell morphology under a bright field by adopting a 40X objective lens, photographing, switching on a mercury lamp to an optical filter corresponding to the excitation wavelength and the emission wavelength of the orange G carbon point material, turning off an external light source to observe the cells which show fluorescence after the orange G carbon point solution is dyed in a dark environment, and comparing after photographing.
As shown in FIG. 3, a comparison of the cell images of the cells under test and normal cells after staining. Wherein FIG. 3a1 shows a cell morphology of a test cell under bright field, and FIG. 3a2 shows a cell morphology of a test cell under fluorescent conditions; FIG. 3b1 shows a cell morphology of a normal cell under bright field, and FIG. 3b2 shows a cell morphology of a normal cell under fluorescent conditions.
As can be seen from FIG. 3, after the orange G carbon dot solution is adopted to dye cancer cells and normal cells, the increase of the cell nucleus volume of the cancer cells can be seen through microscopic observation, the number of cell nuclei is 2-4, the cell morphology is inconsistent, and the cell nuclei of the normal cells are not obvious and the size and the morphology are consistent; and cancer cells proliferate quickly, and the nucleolus is obvious, whereas that of normal cells is not obvious.
Therefore, after the orange G carbon dot solution is used for dyeing the detected cells and the normal cells, the cell morphology is observed and compared by a fluorescence microscope, so that whether the detected cells are cancerous or not can be judged, and the detection method is simple to operate.
The staining reagent for distinguishing normal cells from cancer cells provided by the invention can be applied to the detection of liver cancer cells in example 2 and is also suitable for the detection of other cancer cells.
The embodiments of the present invention have been described in detail above with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention.
Claims (4)
1. A staining reagent for distinguishing normal cells from cancer cells, characterized by: preparing a carbon dot solution by taking orange G as a carbon source, wherein the mass concentration of the orange G is 0.125%, the fluorescence excitation wavelength is 517nm, and the emission wavelength is 304nm;
the dyeing reagent is prepared according to the following steps:
(1) PBS buffer solution is used as electrolyte to prepare an orange G solution with the concentration of 0.002 mol/L;
(2) Electrolyzing the orange G solution serving as a carbon source, and when the solution gradually changes from non-fluorescence to yellow-green fluorescence, preparing the orange G carbon dot solution; the voltage of the electrolysis process is 10V, and the electrolysis time is 8min.
2. A staining reagent for distinguishing between normal and cancerous cells according to claim 1, wherein: the PBS buffer ph=7.
3. A method for detecting cancer cells, characterized in that: the method comprises the following steps:
(1) Staining of the examined cells: taking 100 mu L of detected cells, adding 1mL of PBS solution for rinsing, centrifuging at 3000rpm for 5min, discarding supernatant, replacing the new PBS solution to a volume of 1mL, absorbing 20-50 mu L of orange G with a mass concentration of 0.125%, adding the PBS solution into the dyeing reagent for distinguishing normal cells and cancer cells according to claim 1, sucking the solution after incubating for 10min, and sucking the solution after rinsing by the PBS solution to obtain detected cell liquid;
(2) Normal cell staining: adding normal cells of the same tissue into a centrifuge tube filled with PBS buffer solution, centrifuging, discarding supernatant, absorbing 20-50 mu L of the staining reagent, placing into a centrifuged cell tube, incubating for 10min, absorbing, rinsing with PBS solution, discarding upper liquid, and reserving normal cell solution with suspended bottom;
(3) Dropping 100 mu L of examined cell liquid on the glass slide, then reversely buckling the stained examined cell glass slide on the glass slide, and placing the glass slide under a fluorescence microscope to observe cell imaging; simultaneously sucking 100 mu L of normal cell solution after dyeing treatment to drop on a glass slide, and placing the glass slide under a fluorescence microscope to observe cell imaging; and judging whether the detected cells are cancerous or not through the imaging comparison of the two groups of cells.
4. A method according to claim 3, wherein the PBS solution has a ph=7.2-7.4.
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CN103232029A (en) * | 2012-12-25 | 2013-08-07 | 首都医科大学 | Preparation method and application of green fluorescent carbon dots |
CN103436257A (en) * | 2013-08-27 | 2013-12-11 | 湖南师范大学 | Method for preparing fluorescent carbon dots (C-dots) through electrochemical carbonization of ketone |
CN104787742A (en) * | 2014-01-16 | 2015-07-22 | 中国药科大学 | Method for preparing fluorescent carbon nanoparticles by spontaneous reaction |
CN105316697A (en) * | 2015-12-09 | 2016-02-10 | 青岛大学 | Preparation method for solid-state carbon quantum dot |
CN107254309A (en) * | 2017-07-01 | 2017-10-17 | 中国科学院兰州化学物理研究所 | A kind of preparation method of carbon quantum dot |
CN110982513A (en) * | 2019-11-29 | 2020-04-10 | 郑州大学 | Preparation method of fluorescent carbon dots and application of fluorescent carbon dots in cell imaging |
WO2021087646A1 (en) * | 2019-11-04 | 2021-05-14 | Beijing Normal University | Carbon quantum dots and uses thereof |
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2021
- 2021-07-20 CN CN202110815980.0A patent/CN113466015B/en active Active
Patent Citations (7)
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CN103232029A (en) * | 2012-12-25 | 2013-08-07 | 首都医科大学 | Preparation method and application of green fluorescent carbon dots |
CN103436257A (en) * | 2013-08-27 | 2013-12-11 | 湖南师范大学 | Method for preparing fluorescent carbon dots (C-dots) through electrochemical carbonization of ketone |
CN104787742A (en) * | 2014-01-16 | 2015-07-22 | 中国药科大学 | Method for preparing fluorescent carbon nanoparticles by spontaneous reaction |
CN105316697A (en) * | 2015-12-09 | 2016-02-10 | 青岛大学 | Preparation method for solid-state carbon quantum dot |
CN107254309A (en) * | 2017-07-01 | 2017-10-17 | 中国科学院兰州化学物理研究所 | A kind of preparation method of carbon quantum dot |
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CN110982513A (en) * | 2019-11-29 | 2020-04-10 | 郑州大学 | Preparation method of fluorescent carbon dots and application of fluorescent carbon dots in cell imaging |
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