CN109650387A - A kind of preparation method and its usage of the graphene oxide probe with high-efficiency biomembrane penetration capacity - Google Patents
A kind of preparation method and its usage of the graphene oxide probe with high-efficiency biomembrane penetration capacity Download PDFInfo
- Publication number
- CN109650387A CN109650387A CN201910071429.2A CN201910071429A CN109650387A CN 109650387 A CN109650387 A CN 109650387A CN 201910071429 A CN201910071429 A CN 201910071429A CN 109650387 A CN109650387 A CN 109650387A
- Authority
- CN
- China
- Prior art keywords
- graphene oxide
- probe
- dna
- penetration capacity
- rna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- C01B32/182—Graphene
- C01B32/198—Graphene oxide
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B2204/00—Structure or properties of graphene
- C01B2204/20—Graphene characterized by its properties
- C01B2204/22—Electronic properties
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
- C01B2204/00—Structure or properties of graphene
- C01B2204/20—Graphene characterized by its properties
- C01B2204/32—Size or surface area
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Pathology (AREA)
- Materials Engineering (AREA)
- Nanotechnology (AREA)
- Inorganic Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of preparation method and its usages of graphene oxide probe with high-efficiency biomembrane penetration capacity, it is using conductive black particle as starting material, it first passes around nitric acid oxidation and obtains the carboxylic graphene oxide in surface, then chloride intermediate is obtained with thionyl chloride back flow reaction, again with excessive diamine reactant, obtain the graphene oxide that amino is contained on surface, finally, positively charged cationic compound is modified to surface of graphene oxide by amidation process, obtain the positively charged graphene oxide in surface, the i.e. described graphene oxide probe with high-efficiency biomembrane penetration capacity.Graphene oxide probe of the present invention being capable of penetrating cell film, nuclear membrane, enter nucleus, and can be entered in living body biological gonadal cell across biological living envelope barrier layer by layer, realize the sync mark and image analysing computer to nucleic acid structure on cell and biological living level.
Description
Technical field
The present invention relates to stannic oxide/graphene nano materials synthesis and biomolecule image analysing computer fields, and in particular to a kind of tool
There is the preparation method and its usage of the graphene oxide probe of high-efficiency biomembrane penetration capacity.
Background technique
Nucleic acid is the large biological molecule compound as made of many nucleotide polymerizations, is one of most basic substance of life.
According to its chemical composition and the difference of nucleotidesequence, DNA (DNA) and ribonucleic acid can be divided into
(RNA).The main matter basis that DNA is storage, replicates and convey hereditary information, RNA rise emphatically in protein building-up process
It acts on.Therefore, nucleic acid serves conclusive in a series of great biological phenomenas such as growth, heredity, variation.In living cells
The dynamic change of middle real-time in-situ observation DNA and RNA structure, to vital movements such as the deep transcription for understanding cell, apoptosis and
The pathogenic mechanisms such as tumour generation, virus infection, hereditary variation are disclosed to be of great significance.
However, since cell includes complicated membrane structure, especially the nuclear membrane knot such as cell membrane and various organelle inner membrances
Structure has the selectivity of height to the exchange of substance inside and outside core, being capable of matter transportation and letter inside and outside strict control nucleus
Breath exchange, to maintain the stability of hereditary information.Therefore, can be used at present intracellular DNA and RNA image probe and few
See.Importantly, successfully being distinguished in living cells and biological living due to the limitation of current DNA and RNA image probe
DNA and RNA, and dynamically observing its structure change in real time is still a challenge.Firstly, in the combination dye of DNA and RNA
In, only a small number of intercellular membrane barriers that can be penetrated layer by layer, and tissue and device can be directed through almost without one kind
Official's barrier.Secondly, most of organic dyestuff has photobleaching, prolonged dynamic observation can not be carried out, and can not hold
By the super-resolution scanning imagery for needing high laser energy.Finally, most of dyestuff cannot synchronize differentiation DNA and RNA at present,
It needs simultaneously using a variety of dyestuffs, or even needs the imaging technique by complex and expensive, such as gating technology, although this can part
Solve the problems, such as, but using also resulting in the conveying of dyestuff and the complication of bio distribution simultaneously while polychromatophilia material.
Summary of the invention
The present invention provides a kind of with high-efficiency biomembrane penetration capacity the problems of for above-mentioned prior art
The preparation method and its usage of graphene oxide probe.Graphene oxide probe of the invention has high-efficiency biomembrane penetration capacity
And nucleic acid efficient identification ability, it efficiently solves living cells existing for existing DNA and RNA image probe and biological living is horizontal
Upper film penetration capacity is poor, easy photobleaching, can not synchronize the technical problems such as detection.
The present invention has the graphene oxide probe of high-efficiency biomembrane penetration capacity, is former for starting with conductive black particle
Material, first passes around nitric acid oxidation and obtains the carboxylic graphene oxide in surface, then obtain acyl chlorides with thionyl chloride back flow reaction
Change intermediate, then with excessive diamine reactant, obtain the graphene oxide that amino is contained on surface, finally, by amidation process general
Positively charged cationic compound is modified to surface of graphene oxide, and the positively charged graphene oxide in surface, i.e., the described tool are obtained
There is the graphene oxide probe of high-efficiency biomembrane penetration capacity.Specifically comprise the following steps:
Step 1: commercialized conductive black particle 0.2g (partial size 30nm) is placed in back flow reaction in the nitric acid of 50mL, 6M
It 24 hours, is cooled to room temperature, is centrifuged after reaction, collect supernatant；It is molten that gained supernatant evaporates removing at 180-200 DEG C
Agent obtains red brown solid, dries in a vacuum drying oven, obtains the carboxylic graphene oxide in surface；Nitric acid oxidation obtains
Carboxylic-oxidized graphene have uniform small size, particle size < 10nm.
Step 2: excess thionyl chloride is added in the carboxylic graphene oxide in surface obtained to step 1, is then added dropwise
Excess thionyl chloride is distilled off after reaction, obtains among chloride for a few drop anhydrous DMFs, stirring and dissolving, back flow reaction
Body；Thionyl chloride makees solvent in this step, and DMF plays hydrotropy catalytic action.
Step 3: diamines is added into step 2 gained chloride intermediate, and anhydrous tetrahydro furan is added and makees solvent, 60-
80 DEG C reaction 48-72 hours, silicagel column obtains the graphene oxide that amino is contained on surface after purification；The addition of diamines in this step
Amount is 1.5-2 times of equivalent of chloride intermediate.
In step 3, the diamines includes that ethylenediamine, p-phenylenediamine, hexamethylene diamine etc. can be modified onto carboxylic-oxidized graphene
Diamine compound.
In step 3, eluant, eluent when silica gel column purification is methylene chloride: methanol=5:1-10:1 (v/v).
Step 4: adding excess thionyl chloride into positively charged cationic compound, a few drop DMF are then added dropwise, stir molten
Solution, is back to fully reacting, excessive thionyl chloride is distilled off, then by the positively charged cationization of the chloride of acquisition
The surface that conjunction object is added to step 3 acquisition is contained in the graphene oxide of amino, and anhydrous tetrahydro furan is added and makees solvent, 60-
80 DEG C reaction 48-72 hours, products therefrom is dissolved with ethyl alcohol, revolving, then with ultrapure water dissolution penetrate energy to get the high performance membrane
Power graphene oxide probe.
In step 4, the dosage of positively charged cationic compound is identical as the additional amount of diamines in step 3, i.e. the two matter
Amount is than being 1:1.
In step 4, the positively charged cationic compound is can be coupled by amidation process containing carboxyl
Ionic compound, such as carboxylic butyltriphenylphosphonium bromide phosphine, carboxyl quaternary ammonium salt.
The present invention has the purposes of the graphene oxide probe of high-efficiency biomembrane penetration capacity, is the nucleus in living cells
It is used in position fixing process as detection reagent.
Further, the graphene oxide probe makes during the apoptotic nueleolus of living cells as detection reagent
With can be realized the synchronous imaging of DNA and RNA in living cells.
Further, the graphene oxide probe can be realized in living cells the same of two kinds of DNA and RNA different nucleic acid
Step distinguishes image checking.
The present invention has the purposes of the graphene oxide probe of high-efficiency biomembrane penetration capacity, is in living cells mitosis
It is used in the process as detection reagent, imaging analysis is carried out to the structure dynamics variation of DNA in living cells and RNA.The oxidation
Graphene probe and the combination of DNA and RNA are secured, will not be diffused into the disappearance of cell membrane during mitosis
In cytoplasm.
The present invention has the purposes of the graphene oxide probe of high-efficiency biomembrane penetration capacity, is the reproduction in biological living
It is used in the position fixing process of gland cell DNA and RNA as detection reagent.The graphene oxide probe can pass through life layer by layer
Object envelope barrier carries out imaging analysis to gonadal cell DNA and RNA in biological living.
The biological living is Caenorhabditis elegans adult living.
In the present invention, by the modification of positively charged cationic chemical object, reach the surface potential of graphene oxide probe
20mV or more.Its fluorescence emission spectrum increases red shift with excitation wavelength.
Graphene oxide probe of the present invention can effectively pass through cell membrane and nuclear membrane, synchronous with RNA to intracellular DNA
Imaging.Graphene oxide probe of the present invention has different combinations from double-stranded DNA and single stranded RNA, emits respectively in conjunction with rear
Green and red fluorescence out.
Graphene oxide probe of the present invention can pass through membranes barriers layer by layer, to the DNA in the biological livings such as nematode
It is imaged with RNA.
The invention proposes a kind of cationic surface modification strategies, in the carboxylic fourth that the surface modification of graphene oxide is positively charged
Base triphenylphosphinebromide is successfully realized the transformation of surface of graphene oxide charge from negative to positive.Pass through ammonia with currently used
Base, which ionizes, to be formed positive charged ions and compares, and this direct charged ion is not influenced by environment pH value, and with electronegative DNA
There is higher affinity with RNA.Compared with current detection of nucleic acids probe, which being capable of penetrating cell film and nucleus
Intracellular DNA and RNA is imaged in film.And since probe is different from the combination of double-stranded DNA and single stranded RNA, lead
Cause its in conjunction with DNA and RNA after respectively issue green and red fluorescence, be successfully realized the synchronization zone of intracellular DNA and RNA
It is divided into picture.Currently, this can synchronize the intracellular DNA and RNA of detection with single probe, and biomembrane screen layer by layer can be penetrated
Barrier is realized and is had not been reported to the graphene oxide probe of DNA and RNA in organism living.
Compared with prior art, the beneficial effects of the present invention are embodied in:
1, graphene oxide used in the present invention is that the conductive black particle for removing 30nm by nitric acid oxidation is got, gained oxygen
Graphite alkene just has several nanometers of single-size without the processing such as ultrasound, and this small size is conducive to its penetrating cell core
Film.
2, graphene oxide probe of the present invention the positively charged positive electricity cationic compound from modification, at present often
The method ratio of positive charge is ionized and generated by amino, and the probe of our method synthesis is not by the shadow of extraneous acid or alkali environment
It rings, the affinity of probe and nucleic acid is stronger, it is easier to penetrate biofilm structure layer by layer.
3, graphene oxide probe of the present invention is different from the combination of DNA and RNA, causes different fluorescence, Ke Yitong
Step detection cell and the intracorporal DNA and RNA of biology.Relative to passing through the mixed of the dyeing of a variety of dyestuffs substep or a variety of dyestuffs at present
Conjunction liquid carrys out simultaneously dyeing and compares, our probe application method is simple, and testing result is reliable, can effectively avoid a variety of probes biologies
It is distributed inconsistent the problem of bringing.
4, graphene oxide probe of the present invention, compared with current commercial dyes probe, with strong photostability and low
Cytotoxicity, the in situ imaging detection of long time-histories can be carried out to cell and biological living, can observe in living cells DNA with
The dynamic change of RNA, and can be used for the microscopical 3D imaging of super-resolution (STED).
Detailed description of the invention
Fig. 1 present invention gained high performance membrane penetrates the structural schematic diagram of graphene oxide probe, surface potential variation figure and thoroughly
Penetrate electron microscope.It can be seen from the figure that original graphene oxide contains carboxyl, surface potential is -10mV or so, works as modification
Its potential switchs to positive 7mV after upper p-phenylenediamine, is coupled upper 4 carboxylic butyltriphenylphosphonium bromide phosphine of cationic compound when further
Afterwards, the surface potential of graphene oxide reaches positive 20mV.The differentiation of surface potential has absolutely proved that surface of graphene oxide is repaired
Group is adornd to the decisive action of surface potential.From transmission electron microscope picture as can be seen that the graphene oxide probe has uniform ruler
It is very little, average grain diameter 2-3nm.
Fig. 2 present invention gained high performance membrane penetrates the fluorescence emission spectrum of graphene oxide probe and to double-stranded DNA and single-stranded
The fluorescence spectrum of RNA response.
Fig. 3 present invention gained high performance membrane penetrates imaging of the graphene oxide probe to different cell line cell core DNA.
Fig. 4 present invention gained high performance membrane penetrates the mode of action schematic diagram of graphene oxide probe Yu intracellular DNA and RNA
And its synchronous imaging to DNA in living cells and RNA.
Fig. 5 present invention gained high performance membrane penetrates graphene oxide probe to cell mitogen different times DNA and RNA
Imaging analysis.
It is super to the 3D of living cells mitosis prophase chromosome that Fig. 6 present invention gained high performance membrane penetrates graphene oxide probe
High-resolution (STED) is imaged and the 3D STED of kernel is imaged.
Fig. 7 present invention gained high performance membrane penetrates graphene oxide probe to DNA and RNA in nematode gonadal cell living
Synchronous imaging.
Specific embodiment
Following embodiments are further explanations to summary of the invention using as the explaination to the technology of the present invention content, but this hair
Bright substantive content is not limited in described in following embodiments, those skilled in the art can with and also should know any
Simple conversion or replacement based on invention connotation should belong to protection scope of the presently claimed invention.
Embodiment 1:
1, the conductive black (vxc-72) of 0.2g is added in 100mL round-bottomed flask, is added with stirring the nitre of 50mL 6M
Acid back flow reaction 24 hours, is cooled to room temperature, and centrifugation retains the supernatant of rufous；Supernatant is distilled at 180 DEG C, is obtained
To red brown solid, and further it is put into vacuum drying oven dry.
2,10mL thionyl chloride, stirring and dissolving is added in the solid to step 1 after dry, then 100 μ L anhydrous DMFs is added dropwise,
80 DEG C of back flow reaction 12h, are distilled off excessive thionyl chloride；Then 20mg p-phenylenediamine is added, 2mL anhydrous tetrahydro furan is done
Solvent is stirred to react 72 hours at 60 DEG C, and be cooled to room temperature, obtained solid methylene chloride: methanol=10:1 (v/v) elutes
Agent is purified, and the solid of purifying is dry in vacuum drying oven, spare.
3,4- carboxylic butyltriphenylphosphonium bromide phosphine 20mg adds 10mL thionyl chloride, stirring and dissolving, then that 100 μ L are added dropwise is anhydrous
DMF, 80 DEG C of back flow reaction 12h, is distilled off excessive thionyl chloride；The chloride 4- carboxylic butyltriphenylphosphonium bromide that will be activated
Phosphine, which is added to, to be purified in step 2 in dry solid, and 2mL anhydrous tetrahydro furan is added to make solvent, is stirred to react at 60 DEG C 72 hours,
It is cooled to room temperature；20mL ethyl alcohol is added in obtained solid, then ultrasonic dissolution rotates, and obtained solid ultrapure water dissolves to get arriving
Graphene oxide probe with high-efficiency biomembrane penetration capacity.
Embodiment 2: the nucleus DNA image checking of variety classes cell.
In conjunction with Fig. 3, graphene oxide probe is used for the image checking of cell nuclear dna.Cell kind is in the burnt culture of copolymerization
It on ware, cultivates 24 hours, then probe is added in culture medium, mix, the final concentration of 30 μ g/mL of probe, in cell culture
It is co-cultured 2 hours in case with cell, then carries out fluorescence imaging using laser confocal microscope.Under 488nm excitation, discovery
The nucleus of 12 kinds of cells including tumour cell and normal cell all issues strong green fluorescence, and cytoplasm does not almost have
Have green fluorescence, which show graphene oxide probe can penetrating cell film, nuclear membrane, then with endonuclear DNA
In conjunction with, 488nm excitation under issue green fluorescence.
Embodiment 3: the synchronous imaging of DNA in living cells and RNA is detected.
In conjunction with Fig. 2 and Fig. 4, since graphene oxide probe is different from the combination of DNA and RNA, to cause difference
Fluorescence spectrum response.For double-stranded DNA, graphene oxide probe is embedded into the groove of DNA, causes the enhancing of fluorescence；And
For single stranded RNA, since it does not have rule double-spiral structure as DNA, surface negative charge causes graphene oxide probe
Aggregation reunite, generate red fluorescence.Therefore, after we co-culture probe and cell, swashed respectively with 488nm and 543nm
It shines and co-focusing imaging is carried out to cell, as a result as shown in figure 4, the green fluorescence of 488nm excitation is concentrated mainly on nucleus, and
The red fluorescence of 543nm excitation is concentrated mainly on cytoplasm and kernel.So being realized by graphene oxide probe thin to work
DNA intracellular detection synchronous with RNA's.
Embodiment 4: to the dynamic imaging of cell mitogen different times DNA and RNA.
In conjunction with Fig. 5, the cell synchronization of different division stages is obtained using starvation method.It takes thin in logarithmic growth phase
Born of the same parents after PBS buffer solution cleaning, continue overnight incubation with the culture medium containing 1% serum, then remove and be added with regard to culture medium containing 10%
The culture medium of serum continues to cultivate, and cultivates the different time to obtain the cell in mitosis different times.Then it is added
Graphene oxide probe, respectively under 488nm and 543nm laser, the structure of observation mitosis different times DNA and RNA becomes
Change.From, it can be found that cell membrane disappears during cell mitogen, chromatin high degree of spiralization forms chromosome in Fig. 5,
Under 488nm excitation, chromosome still shows very strong green fluorescence, illustrates graphene oxide probe still in conjunction with chromosome
Together, without because nuclear membrane disappearance and be discharged into cytoplasm, it was demonstrated that affinity strong between probe and DNA.With having
The completion of silk division, probe are also uniformly distributed in two daughter cells.It is intracytoplasmic meanwhile under 543nm excitation
Strong red fluorescence is presented in RNA, and kernel disappears during mitosis, with the completion of division, in two new daughter cells
In, kernel occurs again.
Embodiment 5: the in-situ synchronization image checking of nematode gonadal cell DNA and RNA.
Wild type nematode Bristol N2 is cultivated according to standardization program, i.e., at 20 °C, nematode culture is existed
On nematode growth medium agar plate, using OP50 Escherichia coli as food.The nematode for taking the adult stage, with ultrapure water by nematode adult
It is rinsed from agar plate, is then transferred into the M9 buffer containing graphene oxide probe and cultivates 4 hours, then by line
Worm, which draws out, drips on glass slide, is anaesthetized with NaN3 solution, then covered, and co-focusing imaging is carried out.Excitation
Wavelength is respectively 488nm and 543nm, as a result as shown in Figure 7.
Claims (10)
1. a kind of preparation method of the graphene oxide probe with high-efficiency biomembrane penetration capacity, it is characterised in that:
It is to first pass around nitric acid oxidation using conductive black particle as starting material and obtain the carboxylic graphene oxide in surface, so
Obtain chloride intermediate with thionyl chloride back flow reaction afterwards, then with excessive diamine reactant, obtain the oxidation that amino is contained on surface
Graphene obtains surface finally, modifying positively charged cationic compound to surface of graphene oxide by amidation process
Positively charged graphene oxide, i.e., the described graphene oxide probe with high-efficiency biomembrane penetration capacity.
2. preparation method according to claim 1, it is characterised in that include the following steps:
Step 1: commercialized conductive black particle 0.2g is placed in back flow reaction 24 hours in the nitric acid of 50mL, 6M, reaction knot
It is cooled to room temperature, is centrifuged after beam, collect supernatant；Gained supernatant evaporation of solvent at 180-200 DEG C, obtains rufous
Solid is dried in a vacuum drying oven, obtains the carboxylic graphene oxide in surface；
Step 2: excess thionyl chloride is added in the carboxylic graphene oxide in surface obtained to step 1, several drops are then added dropwise
Excess thionyl chloride is distilled off in anhydrous DMF, stirring and dissolving, back flow reaction after reaction, obtains chloride intermediate；
Step 3: diamines is added into step 2 gained chloride intermediate, and anhydrous tetrahydro furan is added and makees solvent, 60-80 DEG C
Reaction 48-72 hours, silicagel column obtains the graphene oxide that amino is contained on surface after purification；
Step 4: adding excess thionyl chloride into positively charged cationic compound, a few drop DMF are then added dropwise, stirring and dissolving is returned
It is flow to fully reacting, excessive thionyl chloride is distilled off, then by the positively charged cationic compound of the chloride of acquisition
The surface for being added to step 3 acquisition is contained in the graphene oxide of amino, and anhydrous tetrahydro furan is added and makees solvent, and 60-80 DEG C
Reaction 48-72 hours, products therefrom is dissolved with ethyl alcohol, revolving, then is dissolved with ultrapure water to get the high performance membrane penetration capacity oxygen
Graphite alkene probe.
3. preparation method according to claim 2, it is characterised in that:
In step 3, the diamines includes that ethylenediamine, p-phenylenediamine, hexamethylene diamine etc. can be modified to two on carboxylic-oxidized graphene
Amine compounds；The additive amount of diamines is 1.5-2 times of equivalent of chloride intermediate.
4. preparation method according to claim 2, it is characterised in that:
In step 4, the positively charged cationic compound is the ion that can be coupled by amidation process containing carboxyl
Type compound；The mass ratio of diamines is 1:1 in the dosage and step 3 of positively charged cationic compound.
5. a kind of use of any graphene oxide probe with high-efficiency biomembrane penetration capacity prepared in claim 1-4
On the way, it is characterised in that: be to be used during the apoptotic nueleolus of living cells as detection reagent.
6. purposes according to claim 5, it is characterised in that:
The graphene oxide probe uses during the apoptotic nueleolus of living cells as detection reagent, can be realized living thin
The synchronous imaging of DNA and RNA intracellular.
7. purposes according to claim 6, it is characterised in that:
The graphene oxide probe can be realized in living cells two kinds of DNA and RNA different nucleic acid synchronize be distinguished into as detection.
8. a kind of use of any graphene oxide probe with high-efficiency biomembrane penetration capacity prepared in claim 1-4
On the way, it is characterised in that: be to be used during living cells mitosis as detection reagent, to the knot of DNA in living cells and RNA
Structure dynamic change carries out imaging analysis.
9. a kind of use of any graphene oxide probe with high-efficiency biomembrane penetration capacity prepared in claim 1-4
On the way, it is characterised in that: used in the position fixing process of gonadal cell DNA and RNA as detection reagent in biological living.
10. purposes according to claim 9, it is characterised in that:
The biological living is Caenorhabditis elegans adult living.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910071429.2A CN109650387B (en) | 2019-01-25 | 2019-01-25 | Preparation method and application of graphene oxide probe with efficient biological membrane penetration capacity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910071429.2A CN109650387B (en) | 2019-01-25 | 2019-01-25 | Preparation method and application of graphene oxide probe with efficient biological membrane penetration capacity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109650387A true CN109650387A (en) | 2019-04-19 |
| CN109650387B CN109650387B (en) | 2022-04-26 |
Family
ID=66121160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910071429.2A Active CN109650387B (en) | 2019-01-25 | 2019-01-25 | Preparation method and application of graphene oxide probe with efficient biological membrane penetration capacity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109650387B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012800A (en) * | 2019-12-30 | 2020-04-17 | 国家纳米科学中心 | Carbon nanoparticles for scavenging free radicals, and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884430A (en) * | 2006-06-30 | 2006-12-27 | 上海师范大学 | Fluorescent carbon nanometer tube and its preparation method and application |
| US20150080251A1 (en) * | 2012-04-24 | 2015-03-19 | Snu R&Db Foundation | Composition for detecting nucleic acid and method for detecting nucleic acid using same |
| CN104694116A (en) * | 2013-12-06 | 2015-06-10 | 中国科学院大连化学物理研究所 | A carbon-base fluorescence probe for a targeting living cell mitochondrion and preparation thereof |
| CN105218556A (en) * | 2015-09-25 | 2016-01-06 | 江苏大学 | Porphyrin-redox graphene nonlinear optical material and preparation method thereof |
| CN106932368A (en) * | 2017-01-20 | 2017-07-07 | 北京理工大学 | ONOO in a kind of carbon quantum dot targeting detection mitochondria‑Method |
| CN107902642A (en) * | 2017-10-12 | 2018-04-13 | 兰州大学 | A kind of preparation and application study of the graphene quantum dot being imaged for nuclei images and mitochondria |
-
2019
- 2019-01-25 CN CN201910071429.2A patent/CN109650387B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884430A (en) * | 2006-06-30 | 2006-12-27 | 上海师范大学 | Fluorescent carbon nanometer tube and its preparation method and application |
| US20150080251A1 (en) * | 2012-04-24 | 2015-03-19 | Snu R&Db Foundation | Composition for detecting nucleic acid and method for detecting nucleic acid using same |
| CN104694116A (en) * | 2013-12-06 | 2015-06-10 | 中国科学院大连化学物理研究所 | A carbon-base fluorescence probe for a targeting living cell mitochondrion and preparation thereof |
| CN105218556A (en) * | 2015-09-25 | 2016-01-06 | 江苏大学 | Porphyrin-redox graphene nonlinear optical material and preparation method thereof |
| CN106932368A (en) * | 2017-01-20 | 2017-07-07 | 北京理工大学 | ONOO in a kind of carbon quantum dot targeting detection mitochondria‑Method |
| CN107902642A (en) * | 2017-10-12 | 2018-04-13 | 兰州大学 | A kind of preparation and application study of the graphene quantum dot being imaged for nuclei images and mitochondria |
Non-Patent Citations (3)
| Title |
|---|
| XIAOJUAN WANG ET AL.: ""Steering graphene quantum dots in living cells: lighting up the nucleolus"", 《JOURNAL OF MATERIALS CHEMISTRY B》 * |
| YONGQIANG DONG ET AL.: ""One-step and high yield simultaneous preparation of single- and multi-layer graphene quantum dots from CX-72 carbon black"", 《JOURNAL OF MATERIALS CHEMISTRY》 * |
| 胡先运等: ""碳点-核酸/氧化石墨烯荧光探针用于miRNA-21的细胞成像"", 《影像科学与光化学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012800A (en) * | 2019-12-30 | 2020-04-17 | 国家纳米科学中心 | Carbon nanoparticles for scavenging free radicals, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109650387B (en) | 2022-04-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | In situ simultaneous monitoring of ATP and GTP using a graphene oxide nanosheet–based sensing platform in living cells | |
| CN107677656B (en) | A kind of ratio fluorescent nano probe and its application | |
| Jia et al. | A ratiometric fluorescent nano-probe for rapid and specific detection of tetracycline residues based on a dye-doped functionalized nanoscaled metal–organic framework | |
| Huang et al. | Carbon dots for intracellular pH sensing with fluorescence lifetime imaging microscopy | |
| CN105044171B (en) | A kind of preparation method and application of nanometer of platinum dopant/enzyme modification carbon paste electrode | |
| CN105295909A (en) | Method for using phenylenediamine and citric acid for preparing carbon quantum dot marking probe for cell developing | |
| CN104777206B (en) | A kind of fit electrode for detecting terramycin and preparation method thereof | |
| CN106243170B (en) | The synthesis and application of beta galactosidase sensor with aggregation inducing Fluorescence Increasing characteristic | |
| CN109021971B (en) | Fluorescent carbon dot for cell nucleus staining and application and method thereof in cell nucleus imaging | |
| Resa et al. | New dual fluorescent probe for simultaneous biothiol and phosphate bioimaging | |
| Wang et al. | Ferricyanide stimulated cathodic photoelectrochemistry of flower-like bismuth oxyiodide under ambient air: A general strategy for robust bioanalysis | |
| Ripoll et al. | A quantum dot-based FLIM glucose nanosensor | |
| CN104316585B (en) | Combination electrode for NADH electrochemical detection and preparation method of combination electrode | |
| CN109650387A (en) | A kind of preparation method and its usage of the graphene oxide probe with high-efficiency biomembrane penetration capacity | |
| CN105527277B (en) | A kind of preparation method and application of estrone molecule trace electrochemical luminescence sensor | |
| Wei et al. | Self-assembled electroactive MOF–magnetic dispersible aptasensor enables ultrasensitive microcystin-LR detection in eutrophic water | |
| CN105259349B (en) | A kind of preparation for exempting to fix bio-sensing electrode and its application in label-free homogeneous photic electrification learns to farm residual detection and cancer diagnosis | |
| CN111349101B (en) | Turn-on type fluorescent molecular probe responding to biological cell viscosity and preparation method and application thereof | |
| CN113466111A (en) | Single cell analysis system and method and application | |
| CN108426867A (en) | Fe is detected in water3+With the MOF-Cd probes of antibiotic Ceftriaxone Sodium and its preparation method and application | |
| CN106198503B (en) | A kind of electrochemiluminescence sandwich biosensor and preparation and application | |
| Wen et al. | A fluorescent tracer based on castor oil for monitoring the mass transfer of fatliquoring agent in leather | |
| He et al. | Recent advances in electrochemiluminescence-based single-cell analysis | |
| CN111072011B (en) | Preparation of mitochondria-nucleolus reversible migration fluorescent carbon dot and application of mitochondria-nucleolus reversible migration fluorescent carbon dot in monitoring cell activity | |
| Yu et al. | Dynamic layer-by-layer films on nanofiber membrane: a platform for ultra-sensitive bacterial concentration detection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |