CN106967425A - Using cigarette stalk as the preparation method and application of the fluorescent carbon quantum dot of raw material - Google Patents
Using cigarette stalk as the preparation method and application of the fluorescent carbon quantum dot of raw material Download PDFInfo
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- CN106967425A CN106967425A CN201611038893.4A CN201611038893A CN106967425A CN 106967425 A CN106967425 A CN 106967425A CN 201611038893 A CN201611038893 A CN 201611038893A CN 106967425 A CN106967425 A CN 106967425A
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- C09K11/65—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing carbon
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- 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
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- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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Abstract
The invention discloses a kind of preparation method and application using cigarette stalk as the fluorescent carbon quantum dot of raw material.The present invention uses cigarette stalk for raw material to prepare fluorescent carbon quantum dot, and its preparation method operating procedure is simple, it is not necessary to which it is that can obtain fluorescent carbon quantum dot to handle or modify by surface passivator, the biomass of use is widespread in nature, cheap and easy to get, wide material sources, the feature of environmental protection is good.Target fluorescent carbon quantum dot obtained by the present invention all has good solubility and dispersiveness in aqueous, stablize under light illumination, and yield is higher, with quinine sulfate (quantum yield 54%) for object of reference, the quantum efficiency of gained carbon quantum dot is general 34%.
Description
Technical field
The present invention relates to nano material and biological technical field, especially a kind of fluorescent carbon quantum dot using cigarette stalk as raw material
Preparation method and application.
Background technology
Quantum dot (quantum Dots, QDs) is generally soccer star or spherical, is made up of semi-conducting material, stable
Nano-particle of the diameter in 2~20nm.As a kind of novel semiconductor nano material, quantum dot has many unique nanometers
Property.Carbon quantum dot is similar with various metal quantum points, and carbon quantum dot can emit a brilliant light in the case of illumination, but not
It is same as some metal quantum points and contains high noxious metals, it is larger to human body and environmental hazard.Carbon quantum dot main component is carbon,
Carbon quantum dot is nontoxic, and the harm to environment is small, and cost is also less expensive.
The preparation method of carbon quantum dot mainly includes method and from bottom to top two kinds of method from top to bottom.Top-to-bottom method bag
Include as electron radiation, electrochemical oxidation, laser ablation etc., such method test operating condition is strict, and cost is very high, and typically
Quantum yield is not high.Bottom-to-top method includes burning, hydrothermal carbonization method, microwave method, ultrasonic method etc..The advantage of this method
Be to find cheap carbon source as quantum dot synthesis material, while reducing cost, can realize again sustainable development with
And it is environment-friendly, while nontoxic.Other compositions can be mixed in preparation process simultaneously, and quantum yield is higher.High quantum production
Application of the carbon quantum dot of rate in cell imaging and load prescription face is bigger.Biomass energy is widespread in nature, valency
Lattice are cheap, it is easy to obtain.Tobacco is larger in the extensive cultivated area in Yunnan, and after tobacco leaf maturation harvesting, the cigarette stalk amount left is big, is difficult
Processing.
The content of the invention
The purpose of the present invention is：A kind of preparation method and application using cigarette stalk as the fluorescent carbon quantum dot of raw material is provided, it
Both having extended biomass is used for the application of biological technical field, avoids burning to protect environment again, and set up it is a kind of it is simple to operate,
Device simple, the preparation method of raw material environmental protection.
Present invention also offers as fluorescence probe, analyze described carbon quantum dot and cell imaging for ion medicine
Application.
What the present invention was realized in：Using cigarette stalk as the preparation method of the fluorescent carbon quantum dot of raw material, comprise the following steps：
1) calculate according to the mass fraction, take 1-10 parts of cigarette stalks, in 35-50 DEG C of drying to constant weight, to drying to the cigarette stalk of constant weight
Middle 30-50 parts of water of addition, obtain the tobacco rod added water；
2) the cigarette stalk added water is put into hydrothermal reaction kettle, and 18-48h is reacted in 120-240 DEG C of baking oven, obtain anti-
Answer liquid；
3) by step 2) reaction solution that obtains separates under the conditions of 8000rpm centrifugation 15min with large granular impurity and separates simultaneously
The 24-72h that dialyses is placed in bag filter, a water is changed every 6h, that is, obtains fluorescent carbon quantum dot.
Application of the described fluorescent carbon quantum dot in the life or death identification of gram-positive bacteria.
Application of the described fluorescent carbon quantum dot in the life or death identification of Gram-negative bacteria.
Application of the described fluorescent carbon quantum dot in the life or death identification of saccharomycete.
Application of the described fluorescent carbon quantum dot in the life or death identification of filamentous fungi.
Application of the described fluorescent carbon quantum dot in cell imaging.
By adopting the above-described technical solution, compared with prior art, the present invention uses cigarette stalk for raw material to prepare
Fluorescent carbon quantum dot, its preparation method operating procedure is simple, it is not necessary to can obtain by surface passivator processing or modification glimmering
Light carbon quantum dot, the biomass of use is widespread in nature, cheap and easy to get, wide material sources, and the feature of environmental protection is good.Institute of the present invention
Obtained target fluorescent carbon quantum dot all has good solubility and dispersiveness in aqueous, stablizes under light illumination, and
Yield is higher, with quinine sulfate (quantum yield 54%) for object of reference, and the quantum efficiency of gained carbon quantum dot is general 34%.
Fig. 1 is the fluorescence pattern and fluorescence imaging figure of carbon fluorescence quantum in embodiment 1；
Fig. 2 is the transmission electron microscope picture of carbon fluorescence quantum in embodiment 1；
Fig. 3 is the grain size distribution of carbon fluorescence quantum in embodiment 1；
Fig. 4 is the infrared spectrogram of carbon fluorescence quantum in embodiment 1；
Fig. 5 be in embodiment 2 carbon fluorescence quantum to lactic acid bacteria life or death coloration result (excitation wavelength is 552), wherein A
Figure is that dead bacterium B figures are viable bacteria；
Fig. 6 be in embodiment 2 carbon fluorescence quantum to lactic acid bacteria life or death coloration result (excitation wavelength is 488), wherein A
Figure is that dead bacterium B figures are viable bacteria；
Fig. 7 be in embodiment 3 carbon fluorescence quantum to Escherichia coli life or death coloration result (excitation wavelength is 552), wherein A
Figure is that dead bacterium B figures are viable bacteria；
Fig. 8 be in embodiment 3 carbon fluorescence quantum to Escherichia coli life or death coloration result (excitation wavelength is 488), wherein A
Figure is that dead bacterium B figures are viable bacteria；
Fig. 9 be in embodiment 5 carbon fluorescence quantum to saccharomycete life or death coloration result (excitation wavelength is 552), wherein A figures
It is viable bacteria for dead bacterium B figures；
Figure 10 be in embodiment 5 carbon fluorescence quantum to saccharomycete life or death coloration result (excitation wavelength is 488), wherein A
Figure is that dead bacterium B figures are viable bacteria；
Figure 11 be in embodiment 4 carbon fluorescence quantum to trichoderma reesei life or death coloration result (excitation wavelength is 552), wherein
A figures are that dead bacterium B figures are viable bacteria；
Figure 12 be in embodiment 4 carbon fluorescence quantum to trichoderma reesei life or death coloration result (excitation wavelength is 488), its
Middle A figures are that dead bacterium B figures are viable bacteria；
Figure 13 be in embodiment 7 carbon fluorescence quantum to A549 cell dyeings result, (excitation wavelength is 552 (A) and 488
(B)。
Embodiment
Embodiments of the invention：
Embodiment 1：Using cigarette stalk as the preparation method of the fluorescent carbon quantum dot of raw material,
1) 10g cigarette stalks are taken, in 40 DEG C of drying to constant weight, 40mL water is added into the tobacco rod of constant weight to drying；
2) the cigarette stalk added water is put into 100ml hydrothermal reaction kettle, and 36h is reacted in 180 DEG C of baking oven, obtain anti-
Answer liquid；
3) by step 2) reaction solution that obtains separates under the conditions of 8000rpm centrifugation 15min with large granular impurity and separates simultaneously
The 48h that dialyses is placed in bag filter, a water is changed every 6h, that is, obtains fluorescent carbon quantum dot, by the fluorescent carbon quantum dot of acquisition
The aqueous solution freezes standby through vacuum freeze drier.Fluorescence spectrum detection, transmission electron microscope observing, particle diameter point are carried out to carbon quantum dot
Analysis and infrared detection, are as a result shown in Fig. 1-4.Fig. 1 fluorescence spectrums show that the carbon point has maximum absorption band, ultraviolet light at 525nm
Lower irradiation sends blue-fluorescence (in Fig. 1), and transmission electricity is competing and particle diameter statistics shows that the carbon quantum dot size is 4nm or so, red
External spectrum shows that the carbon quantum dot contains-OH, C=O ,-CH, C-O and C=C.
In the present embodiment, cigarette stalk uses Yunnan Wenshan Prefecture cigarette stalk, and hydrothermal reaction kettle is high for the high temperature of polytetrafluoroethyllining lining
Press hydrothermal reaction kettle.
Embodiment 2：Application of the fluorescent carbon quantum dot in terms of the life or death identification of gram-positive bacteria, choose lactic acid bacteria this
Gram-positive bacteria overnight incubation in MRS culture mediums is planted, carbon Vertex Coloring is added, observed under laser confocal scanning microscope
Fluorescence, is as a result shown in Fig. 5-6, the lactic acid bacteria put to death when being excited under 488nm inspires green fluorescence, and bacterium living is then without fluorescence；
Excited under 552nm, the lactic acid bacteria of execution inspires red fluorescence, and bacterium living is then without fluorescence.
Embodiment 3：Application of the fluorescent carbon quantum dot in terms of the life or death identification of Gram-negative bacteria, choose Escherichia coli this
Gram-negative bacteria overnight incubation in LB culture mediums is planted, carbon Vertex Coloring is added, observed under laser confocal scanning microscope
Fluorescence.As a result Fig. 7-8 are seen, the Escherichia coli put to death when being excited under 488nm inspire green fluorescence, and bacterium living is then without fluorescence；
Excited under 552nm, the Escherichia coli of execution inspire red fluorescence, and bacterium living is then without fluorescence.
Embodiment 4：Saccharomycete grows in PDA liquid to be stayed overnight, and is added under carbon Vertex Coloring, laser confocal scanning microscope
Observation.As a result see that the saccharomycete put to death when being excited under Fig. 9-10,488nm inspires green fluorescence, viable bacteria silk is then without fluorescence；
Excited under 552nm, the saccharomycete of execution inspires red fluorescence, and viable bacteria silk is then without fluorescence.
Embodiment 5：Application of the fluorescent carbon quantum dot in terms of the life or death identification of filamentous fungi, trichoderma reesei is in PDA plate
Upper growth takes spore to take mycelia after being connected to PDA liquid medium culture three days after 7 days, adds fluorescent carbon quantum dot, laser copolymerization
Burnt flying-spot microscope observation.As a result Figure 11-12 are seen, it is glimmering that the trichoderma reesei mycelia put to death when being excited under 488nm inspires green
Light, viable bacteria silk is then without fluorescence；Excited under 552nm, the trichoderma reesei mycelia of execution inspires red fluorescence, and viable bacteria silk does not have then
Fluorescence.
Embodiment 6：Application of the fluorescent carbon quantum dot in terms of cell imaging：
The fluorescent carbon quantum dot aqueous solution prepared by embodiment 1 is used to mark A549 lung carcinoma cells, as shown in figure 13, cell shape
State is good, it is seen that target carbon quantum dot does not have cytotoxicity, available for viable cell labelling.Figure 13 A are from left to right followed successively by:
The excitated red fluorescence of 552nm and light field.Figure 13 B are from left to right followed successively by：488 excite green fluorescence and light field.Such as figure result
Show that the carbon quantum dot can be dyed to cell, for cell imaging.
Claims (6)
1. a kind of preparation method using cigarette stalk as the fluorescent carbon quantum dot of raw material, it is characterised in that：Comprise the following steps：
1) calculate according to the mass fraction, take 1-10 parts of cigarette stalks, in 35-50 DEG C of drying to constant weight, add to drying into the cigarette stalk of constant weight
Enter 30-50 parts of water, obtain the tobacco rod added water；
2) the cigarette stalk added water is put into hydrothermal reaction kettle, and 18-48h is reacted in 120-240 DEG C of baking oven, reacted
Liquid；
3) by step 2) reaction solution that obtains centrifuges under the conditions of 8000rpm 15min with large granular impurity and separates and be placed in dialysing
Dialyse 24-72h in bag, changes a water every 6h, that is, obtains fluorescent carbon quantum dot.
2. a kind of application of fluorescent carbon quantum dot as claimed in claim 1 in the life or death identification of gram-positive bacteria.
3. a kind of application of fluorescent carbon quantum dot as claimed in claim 1 in the life or death identification of Gram-negative bacteria.
4. a kind of application of fluorescent carbon quantum dot as claimed in claim 1 in the life or death identification of saccharomycete.
5. a kind of application of fluorescent carbon quantum dot as claimed in claim 1 in the life or death identification of filamentous fungi.
6. a kind of application of fluorescent carbon quantum dot as claimed in claim 1 in cell imaging.
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Cited By (4)
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| CN109135738A (en) * | 2018-09-20 | 2019-01-04 | 河南中烟工业有限责任公司 | A kind of N doping carbon dots and its synthetic method and application based on waste tobacco stem |
| CN109929550A (en) * | 2019-04-16 | 2019-06-25 | 山西大学 | A kind of N doping blue-fluorescence carbon quantum dot and its preparation method and application |
| CN111978959A (en) * | 2020-08-22 | 2020-11-24 | 东北农业大学 | Preparation method and application of fluorescent carbon dots capable of rapidly detecting survival state of yeast cells |
| CN113148979A (en) * | 2021-02-05 | 2021-07-23 | 贵州省烟草科学研究院 | Method for preparing carbon dots with different colors under same wavelength by using tobacco waste and application |
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| CN103482609A (en) * | 2013-10-15 | 2014-01-01 | 浙江师范大学 | Preparation method of carbon quantum dots for detecting Fe<3+> in lake water |
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| CN109135738A (en) * | 2018-09-20 | 2019-01-04 | 河南中烟工业有限责任公司 | A kind of N doping carbon dots and its synthetic method and application based on waste tobacco stem |
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| CN113148979A (en) * | 2021-02-05 | 2021-07-23 | 贵州省烟草科学研究院 | Method for preparing carbon dots with different colors under same wavelength by using tobacco waste and application |
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