CN106497563B - A kind of carbon quantum dot nano material and its preparation method and application - Google Patents

A kind of carbon quantum dot nano material and its preparation method and application Download PDF

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CN106497563B
CN106497563B CN201610927994.0A CN201610927994A CN106497563B CN 106497563 B CN106497563 B CN 106497563B CN 201610927994 A CN201610927994 A CN 201610927994A CN 106497563 B CN106497563 B CN 106497563B
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施季森
陈桢雨
夏兵
陈金慧
霍爱玲
王宾
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Nanjing Forestry University
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Abstract

The invention discloses a kind of carbon quantum dot nano materials and its preparation method and application, this method are as follows: take citric acid, mercaptoethylmaine, polyethyleneimine, it is dissolved in ultrapure water, it stirs and evenly mixs, is ultrasonically treated, at 120-170 DEG C, microwave heating treatment, the mixture that reaction has synthesized is dialysed in ultrapure water through bag filter, vacuum rotating drying instrument is dry after the completion, collection material.The present invention is under microwave controllably auxiliary heat condition, by the carbon quantum dot of mercaptoethylmaine tuning citric acid and the more biological functions of polyethyleneimine hydrothermal synthesis, can get has high brightness fluorescent, or the carbon quantum dot of the different materials such as high charge characterization performance, the demand of different biological applications can be directed to, high aqueous phase dispersibility has preferably been provided on the basis of changing synthesis condition, the cell wall fluorescent marker of the plant living cells of low cytotoxicity, or promote the material of animal and plant cells endocytosis and efficient absorption external world solution, or realize the material of the plant intact cell of delivering nucleic acid transfection tool cell wall.

Description

A kind of carbon quantum dot nano material and its preparation method and application
Technical field
The invention belongs to have organic carbon quantum dot synthesis technical field of biological function, it is related to the controllable auxiliary heat of microwave and promotes Mercaptoethylmaine tuning citric acid synthesizes the preparation method of fluorescent polymer or carbon dots with polyethyleneimine, and in particular to a kind of carbon amounts Son point nano material and its preparation method and application.
Background technique
With the rapid development of genome high throughput sequencing technologies, researcher has accumulated the genome sequence of many species Column, the research of these species also enter functional genome's epoch therewith.Wherein, many genes that Bioinformatics Prediction goes out need The research of Function Identification and function interaction is carried out, therefore, a variety of novel, accurately gene functional research technology is constantly gushed It is existing, such as gene editing technology, RNA interference, site-directed point mutation, gene site-directed insertion.These technologies require to construct outer Source nucleic acid molecule is delivered into corresponding intracellular competence exertion effect, and can more importantly obtain transgenic plant.Currently, planting In object transgenic acceptor system, the target of regeneration plant is may be implemented in somatic embryo regenerating system, and in various plants It is middle successfully to construct, it is some to have carried out the factorial production, and the phenotype of the transgenic plant obtained can also pass through plant phenotype Group learns high-throughput character parting and phenotype-genomic data related analysis technology is effectively collected and surveyed.Therefore, top load Amount is stablized, and the Gene transfer techniques of efficient or even a variety of large fragment nucleic acid cotransfections are compeled as each plant species genetic engineering Be essential the key technology asked.But traditional virus, mediated by agriculture bacillus by exogenous genetic material introduced plant cell inner expression Technology, have acceptor material easy to pollute, transfect species narrow range, limitations such as transfected plasmids length is limited, and particle gun are shown Microinjection, electric shocking method, waiting Typical physicals method, there are conversion cost height, low efficiencys, it is difficult to which converting has the plant of cell wall thin The problems such as born of the same parents, cell easy damaged, it is difficult to meet the demand of increasingly urgent plant intact cell high-efficient transgenic.Surface can repair Functional group is adornd, there is multifunctional nano material to have conducted a research extensively in animal as nucleic acid delivery vehicle, studied The advantages that material out has absorption, compression DNA, and DNA is protected not to be digested degradation, and transgene efficiency is high;And nano material With its lesser partial size, surface-active, can be absorbed into cytoplasm by plant cell, by the difference of its structure and surface charge, Can specific localization into different subcellular structures, or even by transportation system in plant, specifically in separate living tissue Middle enrichment.Some nano materials can have launch wavelength narrow range, the excellent fluorescence property such as fluorescent emission intensity height.Cause This, with nano material development and application in the top load of plant, high efficiency nano gene delivery vector is imperative.
Polyethyleneimine (PEI) is a kind of polycation high molecular polymer containing abundant nitrogen-atoms, with 1KDa To the wide molecular weight ranges of 1000KDa, it is divided into linear and branched two kinds of molecular structures, structural framework (- CHI-CHZ-NH-) In every 3 atoms there is an amino, can protonate in physiological conditions, so that PEI positive electricity with higher Lotus density occurs electrically to neutralize, can effectively adsorb, press by the nucleic acid of electrostatic adsorption and negatively charged phosphate group Contract negatively charged nucleic acid molecules, forms stable nanometer-nucleic acid complexes, and surface charge is in neutrality or weakly positive, with surface Negatively charged cell membrane interaction and enter intracellular, and nucleic acid molecules can be protected from degrading in Cytolysosome The degradation of enzyme protonates in acid endosome, adsorbs endosome intermediate ion, changes the inside and outside flow of water, leads to outer ion Interior stream makes endosome swelling fracture, i.e. generation " proton sponge effect ", to release the nucleic acid molecules from PEI separate out Or the compound of PEI and nucleic acid molecules, promote nucleic acid delivery to nucleus, realizes gene stable transfection.By further with its He modifies functional molecular, can obtain higher targeting and transfection efficiency.So polyethyleneimine is thin applied to animal A kind of in-vitro transfection cationic polymer with compared with high transfection efficiency of born of the same parents, be widely used as nucleic acid genomic medicine sun from Sub- polymer carrier system.But the potential cytotoxicity of PEI can be significantly increased with the increase of molecular weight, it is especially flat The PEI that average molecular weight is 25000.Secondly the group of the excessive tool positive charge in the surface PEI and the structure of dispersion, outside being applied to When layer has the delivery of nucleic acids of the plant intact cell of cell wall, cell wall is usually adsorbed only on.Due to these disadvantages In the presence of limiting the application of PEI.But it according to the literature, is synthesized using citric acid with the PEI hydro-thermal reaction of low molecular weight high/super Molecularly Imprinted Polymer is enriched with the positively charged group of PEI on individual molecule, improves plasmid adsorbance, improves transfection efficiency;It will Hydrophilic polymer such as PEG modification reduce positive surface charge, while also reducing toxicity on PEI.It is ground currently, having Study carefully and shows that the PEI for being 25,000 using molecular weight (MW) as efficient protoplast transient expression plasmids delivery vector, is in pH Plasmid DNA is combined under conditions of 7.0 in 50 μ L solution, TEM negative staining detects answering which form the nano-scale of 100-200nm Object is closed, transfection protoplasts of Arabidopsis thaliana broken by ultrasonic average efficiency reaches 65% when N/P ratio is 5.And the polyamide-amide of high positive charge group Type dendrimer (PAMAM) can deliver plasmid and enter in the plant cell with cell wall, realize transgenosis.
Carbon quantum dot (C-dots) is as having unique physico-chemical property, and especially it is with good biocompatibility 4th main group is the nano material of Material synthesis, has been widely applied to bio-sensing, pharmaceutical carrier and fluorescence imaging etc. at present Different field of biomedical research.For other nano materials, C-dots novel has photoluminescent property as a kind of Zero dimension carbon nanomaterial, have the advantage that 1. the relative molecular mass of C-dots and partial size are all small, diameter is generally 1 ~ 10 nm;Carbon is one of most common element of distributed in nature, and constitutes the most important element of all life body on the earth.Example Such as, the skeleton of the basic structural unit amino acid of life entity, protein, nucleotide etc. is all made of carbon, therefore, C- Dots is usually nontoxic or hypotoxicity to life entity, has good biocompatibility and environment friendly;3. having one Member excites the photoluminescent property of polynary transmitting, while also absorbing narrow emission characteristics with excellent width.There is biology in C-dots On the basis of the multiple advantages of materialogy, existing functional macromolecule is combined with organic matter existing for cell itself, or directly The nano materials existing research reports such as the carbon quantum dot with the synthesis small particle such as special cells content such as diatom.
Summary of the invention
Goal of the invention: the deficiencies in the prior art are directed to, the first purpose of the invention is to provide a kind of carbon quantums Point nano material has the features such as transgene efficiency is high, degradable, toxicity is low.A second object of the present invention is to provide one kind The preparation method of above-mentioned carbon quantum dot nano material.Third purpose of the present invention is to provide answering for above-mentioned carbon quantum dot nano material With.
Technical solution: in order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention are as follows:
A kind of preparation method of carbon quantum dot nano material: citric acid, mercaptoethylmaine, polyethyleneimine are taken, is dissolved in ultrapure It in water, stirs and evenly mixs, is ultrasonically treated, at 120-170 DEG C, microwave heating treatment, the mixture that reaction has been synthesized is through dialysing Bag is dialysed in ultrapure water, and vacuum rotating drying instrument is dry after the completion, collection material.
The preparation method of the carbon quantum dot nano material, citric acid and mercaptoethylmaine mass ratio are not less than 1, polyethylene The mass ratio of imines and mercaptoethylmaine is 1-5.
The preparation method of the carbon quantum dot nano material, ultrasonic power is 200 W, frequency is 28 KHz, ultrasound 3 min。
The preparation method of the carbon quantum dot nano material, it is characterised in that: microwave power is 500 W, heating 10 min。
The preparation method of the carbon quantum dot nano material, dialyse 36 h in 45 DEG C of ultrapure water, and 10 h are changed once Ultrapure water.
The preparation method of carbon quantum dot nano material carbon quantum dot nano material obtained.It is positively charged, grain Diameter is small, and the carbon quantum dot with high brightness fluorescent can adsorb negatively charged substance, such as nucleic acid molecules, with suitable proportion In conjunction with formation compound can affine cell wall, and with cell membrane interaction, active cell endocytosis, into intracellular, Realize the delivering of allogenic material.
Application of the carbon quantum dot nano material as cell wall fluorescent marker.
The carbon quantum dot nano material is as the material for promoting animal and plant cells endocytosis and efficient absorption external world solution Application.
Material of the carbon quantum dot nano material as the plant intact cell for realizing delivering nucleic acid transfection tool cell wall The application of material.
The utility model has the advantages that being directed to the deficiencies in the prior art, of the invention its has following advantage:
One, synthetic method is simple and efficient: reaction substrate used in the present invention can dissolve in water, therefore use water Dispersion, dissolution easily adjustment parameter can realize that the tuning of mercaptoethylmaine synthesizes using microwave synthesizer without harsh conditions Function.
Two, product property is tunable: the polymer or carbon quantum dot that the present invention synthesizes have and can issue under ultraviolet lighting The blue and white fluorescence of high brightness, also there is a material for not having fluorescence, but its such as electropositive performance can be relatively more in other respects It is excellent.Material with high brightness fluorescent can be applied to fluorescent visual analysis, and high electropositive material can be used as absorption Carrier, absorption load nucleic acid, drug or nutrients and other items.
Three, shielded surfaces active group, be conducive to cell wall spread, active cell endocytosis: be used only citric acid and sulfydryl The high molecular polymer of ethamine synthesis is compared, and the fluorescence radiation for the material tool high brightness that the present invention obtains, partial size is small and uniform, water Phase dispersion degree is very high, does not form aggregate.And it is not adhere to Outer surfaces of cell wall, it is able to enter disperse in cell wall, can be answered With for cell wall reactive fluorescent dye.After carbon quantum dot saves 2 years in ultrapure water, fluorescent characteristic does not change.
Four, realize biological function it is more: the material that the present invention synthesizes based on have high positive charge cationic polymer, with And there is the active mercaptoethylmaine of potential source biomolecule, and biological glycometabolism intermediate product citric acid, through microwave controllably auxiliary thermal synthesis Compound toxicity it is adjustable very low, and the respective function of reaction substrate can be integrated, as PEI is adsorbed Electronegative substance such as plasmid etc., and obtain the fluorescence radiation of more performances such as high brightness.So the tuning that the present invention is told Synthetic method can optimum synthesis be suitble to the materials of multiple application scenarios, be such as suitable for the high brightness carbon quantum of cell wall imaging Point;The nucleic acid delivery vector that plasmid is adsorbed and delivered with positive charge group stimulates the material etc. of cell endocytic.
Detailed description of the invention
Fig. 1 is synthetic material figure of the embodiment 1 under white light, ultraviolet light;
Fig. 2 is the DLS grain-size graph of 1 synthetic material of embodiment;
Fig. 3 is the Zeta potential figure of 1 synthetic material of embodiment;
Fig. 4 is the uv absorption spectra of 1 synthetic material of embodiment;
Fig. 5 is the fluorescence spectra of 1 synthetic material of embodiment;
Fig. 6 is fluorescence radiation figure of 1 synthetic material of embodiment under Zeiss microscope difference fluorescence channel;
Fig. 7 is that 1 synthetic material of embodiment and Liriodendron Suspension Cells are incubated under rear low power lens for 24 hours (100 ×) altogether and detect Fluorescence distribution figure in cell;
Fig. 8 be 1 synthetic material of embodiment handled in ultrapure water Liriodendron suspension system it is unicellular after cell fluorescence distribution Figure;
Fig. 9 be 1 synthetic material of embodiment handled in PBS Liriodendron suspension system it is unicellular after cell fluorescence distribution map;
Figure 10 be 1 synthetic material of embodiment be incubated for altogether with Liriodendron Suspension Cells the cell UV absorption after 48h with it is glimmering Light spectrogram;A, ultra-violet absorption spectrum;B, ultrapure water blank control group fluorescence spectrum;C, PBS and synthetic material processing group fluorescence Spectrum;D, ultrapure water and synthetic material processing group fluorescence spectrum;
Figure 11 is the streaming fluorescence detection map figure of cell after 1 synthetic material of embodiment and Liriodendron cell absorption 36h;a For the cell map handled through synthetic material, b is untreated cell map;A-1, b-1: λex =355 nm, λem = 410 The cell distribution map of -510 nm Air conduct measurement fluorescence intensities and cellular granularity (SSC);A-2, b-2: λex = 355 nm、 λem =410-510 and λex = 488 nm、λem The cell distribution map of=490-570 nm Air conduct measurement fluorescence intensities;
Figure 12 is that embodiment 2 uses being incubated for altogether after one synthetic material of embodiment absorption plasmid with Liriodendron suspension cell Mediated cell transfection figure after for 24 hours;Realize transient expression;
Figure 13 is that 3 preferred reactant of embodiment matches the fluorescent carbon point synthesized with reaction condition active cell in ultrapure water Endocytosis figure;Structure shown in red arrow is endocytosis vacuolar membrane, and structure shown in green arrow is inside endosome.
Specific embodiment
The present invention is described further combined with specific embodiments below.Following embodiment is merely to illustrate the present invention, But it is not limited to specific range of the invention.
Embodiment 1
By 0.15 g citric acid, 0.13 g mercaptoethylmaine, 0.1 g polyethyleneimine (Mw=800) is dissolved in 3 mL Ultrapure water stirs and evenly mixs, ultrasound (200 W of power, 28 KHz of frequency) 3 min, and 170 DEG C, single mold microwave synthesizer (power 500 W 10 min) are heated, the mixture that has synthesized of reaction is dialysed 36 through bag filter (Mwco:1000) in 45 DEG C of ultrapure water H, 10 h change a ultrapure water, and collection material after vacuum rotating drying instrument is dry is detected glimmering with ultraviolet lamp (305 nm, 365 nm) Light shines, as shown in Figure 1, the brown color that synthetic material is transparent under white light, has at ultraviolet lamp (305 nm, 365 nm) The fluorescence radiation of high brightness.Malvern particle instrument detects the DLS partial size and Zeta potential value of synthetic material, partial size about 0.7 Nm, it is relatively small;Institute's band is positive charge, about+8.18 mV, therefore it can adsorb electronegative plasmid and affine absorption band The cell wall of negative electrical charge, is shown in Fig. 2 and Fig. 3.It can be found using ultraviolet/visible spectrophotometer ultra-violet absorption spectrum as shown in Figure 4 Its UV absorption wave-length coverage is 300-400 nm, and maximal ultraviolet absorption peak is in 350 nm or so.Sepectrophotofluorometer inspection Survey its fluorescence spectrum.It was found that its fluorescence emission wavelengths range is between 400-525 nm, maximum emission wavelength is in 450 nm Left and right, sees Fig. 5.It by the material liquid-transfering gun of 1 μ L, is added dropwise on glass slide, is dried in 60 DEG C of baking ovens, Yu Caisi fluorescence is aobvious Fluorescence radiation is detected under micro mirror fluorescence detection channel.It can in multiple fluorescence detection channels such as FITC(λ ex=455-495 nm, λ em=505-555 nm), CFP(λ ex=426-446 nm, λ em=460-500 nm) in be detected, see Fig. 6.It will system Standby material and the unicellular total incubation of suspension system is all thin in the visual field using low power objective in Zeiss fluorescence microscope The cell wall of born of the same parents all has synthetic material specificity fluorescent, shows its fluorescence intensity height, as shown in Figure 7.
Take the Liriodendron for suspending and cultivating unicellular with 300 mesh cell sieves, 3000 rpm are centrifuged 8 min after dividing equally, suck Clearly, cell culture PBS(pH=7.4 are separately added into) solution and ultrapure water take two kinds of cell suspending liquids after blowing and beating suspension cell Each 2 mL is separately added into 100 μ L synthetic materials, mixes, compares group respectively with the cell suspending liquid of non-plus nano material.23 DEG C, dark culture takes Zeiss fluorescence microscope detection synthetic material adherent cell result after 500 μ L cleaning twice afterwards for 24 hours;Comparison The cell that Fig. 8 and Fig. 9 discovery is suspended using transparent brown color synthetic material processing ultrapure water or PBS, it is molten in total incubation Liquid by transparent yellow, change it is colorless and transparent, after cell cleaning, in conjunction with the cell of synthetic material in pale brown under ultrapure aqueous systems Color, and the brown color of the cell under PBS system in conjunction with synthetic material is shallower, synthetic material may be absorbed by cell degrades.Analysis The fluorescence distribution of cell under ultrapure water control and PBS environment can be after synthetic material and Liriodendron cell are incubated for 24 h altogether It is distributed in entire cell wall, does not contact directly especially in the cell plates of solution postmitotic cell and also detect that fluorescence signal (shown in Fig. 9 orange arrows), show fluorescent carbon point cell wall distribution do not have specificity, and may in cell wall energy Enough spread.Ultraviolet/visible spectrophotometer is used after 48 h, the cell after fluorescence spectrophotometer measurement cleaning twice Ultra-violet absorption spectrum and fluorescence spectrum.As shown in Figure 10, processing is incubated for altogether in ultrapure water and PBS respectively by synthetic material The single celled ultra-violet absorption spectrum of suspension system and fluorescence spectrum, through synthetic material treated ultra-violet absorption spectrum in ultrapure water With untreated more similar map, and cell in PBS through synthetic material treated ultra-violet absorption spectrum and the above two Significantly different, the increase absorption intensity with Detection wavelength is in be gradually reduced trend;The fluorescence spectrum for comparing these three processing groups, can Know, in the material-specific fluorescence wave crest that has of cell and ultrapure water handled in PBS through synthetic material through synthetic material at The material-specific fluorescent emission wave crest of the cell of reason is compared to more significant;Synthetic material still needs specificity in extraneous solution Ion makes cell generate plasmolysis after total incubation 48h, and activates endocytosis, is formed on the inside of the cell membrane separated with wall round Significantly with the endosome of synthetic material specificity fluorescent.
300 mesh cell sieves are carried out in the third day of third time culture to the suspension cell for the culture that suspends every 7 days subculture 2 times Repeatedly sieving obtains the cell for being sized for flow cytomery or screening, is incubated for 36 h altogether with fluorescent carbon point synthetic material Afterwards, it after cleaning 3 times with PBS, suspends carry out the corresponding fluorescence channel detection of flow cytometer again.
On the basis of the visual research that the fluorescence pair using synthetic material interacts with suspension cell, obtained by sieving The suspension system that must be sized for flow cytomery sorting is unicellular, it is used streaming after synthetic material is handled in PBS Cell instrument detection, cellular granularity (SSC) is more concentrated after processing, forms apparent peak type than blank control, the nm of λ ex=355, The sense channel of λ em=410-510 nm be suitble to detect synthetic material fluorescence, altogether be incubated for after cell compared with the control, The number of cells that detection is found to have more high fluorescent under this channel increases relatively, shows synthetic material to the affine absorption of cell Ability it is preferable, have the fluorescence for having the specificity of synthetic material high-intensitive compared with many cells, see Figure 11.
Embodiment 2
The material that 20 μ L embodiments 1 synthesize is dispersed in aqueous systems, is added in ultrapure water with 10 μ g plasmids, is supplied total Amount is 50 μ L, and ice-bath ultrasonic shakes 3min in ultrasonic washing instrument, is placed in dark place and stands in conjunction with after 30 min, is added through 300 Mesh cell sieve be sieved Liriodendron suspension system it is unicellular in, mix, be then transferred into tissue culture plate, add 1 mL ultrapure water, 24 Distribution of the fluorescent carbon point in Liriodendron Suspension Cells is observed after h.Synthetic material has certain positive electricity known to embodiment 1 Lotus group, can by electrostatic interaction adsorb plasmid, is incubated for altogether with cell can adherent cell wall, as shown in figure 12, absorption GFP Egfp expression plasmid can be deliverrf into cell and express, and the green for making entire cytoplasm all there is high brightness is glimmering Light, fluorescence distribution is visibly different after this interacts with exclusive use synthetic material and cell, and synthetic material can deliver Plasmid enters expression in the intact plant with cell wall.
Embodiment 3
Using the different ratio of reaction substrate, synthesis material is reacted through 10 min of microwave synthesizer at a temperature of differential responses Material, by 0.15 g citric acid, 0.08 g mercaptoethylmaine, it is ultrapure to be dissolved in 3 mL for 0.2 g polyethyleneimine (Mw=800) Water stirs and evenly mixs, and 3 min of ultrasound, 150 DEG C, microwave synthesizer heats 10 min, and the mixture that reaction has been synthesized is through dialysing Bag (Mwco:1000) is dialysed 36 h in 45 DEG C of ultrapure water, and 10 h change a ultrapure water, after vacuum rotating drying instrument is dry Collection material, take 200 μ L and 1 mL Liriodendron suspension system it is unicellular carried out in 3 mL ultrapure waters it is total be incubated for 24 h, use fluorescence Material-specific fluorescence finds plasmolysis occur in the cell of the sample treatment of synthesis in microscope detection discovery cell, And there are multiple endosomes with specificity fluorescent being distributed on the inside of cell membrane in multiple cells, it may be in this condition The sample of lower synthesis being capable of activated cell endocytosis.As shown in figure 13.
The present invention is to tune polyethyleneimine and citric acid using mercaptoethylmaine, by microwave-assisted controllable hydrothermal synthesis legal system For carbon quantum dot out, polyethyleneimine is the cationic polymer that molecular weight can be changed in reaction substrate, controls different molecular weight The quality of polyethyleneimine and mercaptoethylmaine consumption is to control reaction temperature (120-170 DEG C) between 1 ~ 5 and react than range Duration (6-10 min) is conducive to that different condition, the hydro-thermal of auxiliary mercaptoethylmaine tuning polyethyleneimine and citric acid is arranged The performance change of synthetic product, can be optimized with high aqueous phase dispersibility, and the cell wall of the plant living cells of low cytotoxicity is glimmering Signal object, or promote the material of animal and plant cells endocytosis and efficient absorption external world solution, or realize that delivering nucleic acid transfection tool is thin The material of the plant intact cell of cell wall.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art Member, without departing from the inventive concept of the premise, can also make several improvement, these improvement also should be regarded as protection of the invention In range.

Claims (8)

1. a kind of preparation method of carbon quantum dot nano material, it is characterised in that: citric acid, mercaptoethylmaine, polyethyleneimine are taken, It is dissolved in ultrapure water, stirs and evenly mixs, be ultrasonically treated, at 120-170 DEG C, microwave heating treatment will react the mixture synthesized It dialyses in ultrapure water through bag filter, vacuum rotating drying instrument is dry after the completion, collection material;Wherein, citric acid and mercapto For base ethamine mass ratio not less than 1, the mass ratio of polyethyleneimine and mercaptoethylmaine is 0.769.
2. the preparation method of carbon quantum dot nano material according to claim 1, it is characterised in that: ultrasonic power 200 W, frequency is 28 KHz, 3 min of ultrasound.
3. the preparation method of carbon quantum dot nano material according to claim 1, it is characterised in that: microwave power is 500 W heats 10 min.
4. the preparation method of carbon quantum dot nano material according to claim 1, it is characterised in that: ultrapure at 45 DEG C Dialyse 36 h in water, and 10 h change a ultrapure water.
5. the preparation method carbon quantum dot nanometer material obtained of the described in any item carbon quantum dot nano materials of claim 1-4 Material.
6. application of the carbon quantum dot nano material as cell wall fluorescent marker described in claim 5.
7. carbon quantum dot nano material described in claim 5 is as promotion animal and plant cells endocytosis and efficient absorption external world solution Material application.
8. carbon quantum dot nano material described in claim 5 is completely thin as the plant for realizing delivering nucleic acid transfection tool cell wall The application of the material of born of the same parents.
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