CN109852379A - The area near-infrared II fluorescence quantum cell membrane tagging system, labeling method and application - Google Patents
The area near-infrared II fluorescence quantum cell membrane tagging system, labeling method and application Download PDFInfo
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- CN109852379A CN109852379A CN201910187615.2A CN201910187615A CN109852379A CN 109852379 A CN109852379 A CN 109852379A CN 201910187615 A CN201910187615 A CN 201910187615A CN 109852379 A CN109852379 A CN 109852379A
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Abstract
The invention discloses a kind of area near-infrared II fluorescence quantum cell membrane tagging system, labeling method and applications.The area near-infrared II fluorescence quantum cell membrane tagging system includes the area near-infrared II fluorescence quantum, and modification is connected to the hydrophilic ligand and hydrophobic ligand on the area near-infrared II fluorescence quantum surface.The labeling method includes: to mix cell to be marked with the area aforementioned near-infrared II fluorescence quantum cell membrane tagging system, form label system, total incubation is carried out later, in the phospholipid bilayer for making the area near-infrared II fluorescence quantum cell membrane tagging system insertion cell membrane, the label to cell membrane is realized.The area near-infrared II fluorescence quantum cell membrane tagging system of the invention can successfully imbed in the phospholipid bilayer of cell membrane, realize label and fluorescence imaging to cell membrane, and the label on active somatic cell has huge application potential and value.
Description
Technical field
The present invention relates to a kind of cell membrane tagging systems, and in particular to fluorescence quantum cell membrane marker system, the area near-infrared II
System, labeling method, and the application for cell membrane with high-affinity label and cell membrane imaging, and can be used as cell
The tool of membrane marker is applied to the purposes in research relevant to cell membrane function, belongs to nano meter biomaterial technology neck
Domain.
Background technique
Cell membrane is to repeat to form as basic unit by Phospholipids bilayer molecules, i.e. phospholipid bilayer, is inlayed thereon
There are various types of memebrane proteins and sugar and glycolipid in conjunction with memebrane protein, is related to various kinds of cell process, such as matter transportation,
Cellular signal transduction, cell adherence, ionic conduction etc..In order to better understand the function of cell membrane with effect and to cell
The influence of vital movement just has to realize the visualization to cell membrane.Currently commercially already present cell membrane label probe
It is concentrated mainly on 400-900nm wave band, such as DiI cell membrane fluorescent red-orange probe, DiI can issue orange red after being excited
Fluorescence, but this kind of fluorescence probe causes its light quantum intensity lower and lifetime of excited state since constant is quenched with very high
It is very short, fluorescence probe is quenched easily, the long-term observation being unfavorable in cell membrane research process；In addition, going back some fluorescence probes
Since its excitation wavelength is in ultraviolet region, excitation energy is higher to cause irradiation for a long time to cause biggish damage to cell,
And it is unfavorable for the progress of research.Meanwhile conventional fluorescent probe (400-900nm) absorbs by force since living tissue has it, is high
The disadvantages of scattering and high spontaneous background fluorescence, faces tissue penetration depths low (< 5 millimeters), space point in living imaging research
The challenge such as resolution low (about 1000 microns) and temporal resolution low (about 1000 milliseconds), seriously restricts its extensive use.
Summary of the invention
For the main object of the present invention aiming at the above status, providing one kind can be to cell membrane highly efficient labeling and long-time
The area the near-infrared II fluorescence quantum cell membrane tagging system of blur-free imaging and corresponding labeling method, to overcome existing cell membrane
The deficiencies of fluorescence labeling probe is easily quenched, penetration depth is shallow and tissue autofluorescence is high.
Another object of the present invention, which also resides in, provides answering for near-infrared II area's fluorescence quantum cell membrane tagging system
With.
For realization aforementioned invention purpose, the technical solution adopted by the present invention includes:
The embodiment of the invention provides a kind of area near-infrared II fluorescence quantum cell membrane tagging systems comprising: it is close red
The outer area II fluorescence quantum, and modification are connected to the hydrophilic ligand on the area near-infrared II fluorescence quantum surface and hydrophobic
Property ligand.
In some embodiments, the hydrophilic ligand includes glutathione, dihydrolipoic acid, thioacetic acid, sulfydryl third
Any one in acid or two or more combinations.
Further, the hydrophobic ligand includes lauryl mercaptan ligand.
The embodiment of the invention also provides the preparation method of the area aforementioned near-infrared II fluorescence quantum cell membrane tagging system,
Comprising: which modifying hydrophobic ligand, hydrophilic ligand respectively is connected to the area near-infrared II fluorescence quantum surface, obtain close red
The outer area II fluorescence quantum cell membrane tagging system.
In some embodiments, the preparation method specifically includes:
So that hydrophobic ligand modification is connected to the area near-infrared II fluorescence quantum surface, forms hydrophobic ligand quantum dot；
Hydrophilic ligand, hydrophobic ligand quantum dot and solvent are uniformly mixed, stirred, is obtained after centrifugation described close red
The outer area II fluorescence quantum cell membrane tagging system.
The embodiment of the invention also provides the area aforementioned near-infrared II fluorescence quantum cell membrane tagging systems in cell membrane mark
Purposes in note or fluorescence imaging field.
Correspondingly, the embodiment of the invention also provides a kind of cell membrane labeling methods comprising: make cell to be marked with
The mixing of the area aforementioned near-infrared II fluorescence quantum cell membrane tagging system, forms label system, carries out total incubation later, makes described
The area near-infrared II fluorescence quantum cell membrane tagging system is embedded in the phospholipid bilayer of cell membrane, realizes the mark to cell membrane
Note.
Relative to existing labeling method and imaging means, the present invention is had the advantage that
1) the double ligand modified II area's fluorescence quantum cell membrane tagging systems of near-infrared of the hydrophilic/hydrophobic that the present invention uses
Good biocompatibility can successfully imbed in the phospholipid bilayer of cell membrane, and toxicity is low, almost to eucaryotic cell structure and function
It does not influence, and cell membrane can be marked to greatest extent and fluorescence imaging；
2) area the near-infrared II fluorescence quantum cell membrane tagging system that the present invention uses has very strong fluorescent stability,
Prolonged continuous imaging may be implemented, avoid the defect that conventional fluorescent dyestuff is easily quenched very much, primary label can be more
It is observed in Long time scale, simplify experimental implementation and can avoid marking cell repeatedly bring structure and function to damage
Evil；
3) biological tissue can be significantly reduced in the area the near-infrared II fluorescence quantum cell membrane tagging system that the present invention uses
Absorption and scattering to photon have better tissue penetration depths and higher spatial resolution, can overcome existing tradition
The defects of membrane marker fluorescence probe is easily quenched, and penetration depth is shallow and tissue autofluorescence is high, can preferably carry out cell membrane
Label and correlation function research, especially have bigger application prospect and value in the label on living animal cell, avoid
The prior art can only be in the defect that cellular level is operated.
Detailed description of the invention
Fig. 1 is to be surface modified in an of the invention exemplary embodiments to the area near-infrared II fluorescence quantum and cell marking
Schematic diagram.
Fig. 2 is GSH-DT-Ag prepared by the embodiment of the present invention 12The transmission electron microscope picture of S quantum dot.
Fig. 3 a- Fig. 3 c is that the embodiment of the present invention 3 marks PC-12 cell, spongiocyte and primary neuronal cell respectively
Fluorescence imaging figure afterwards.
Fig. 4 is hydrophily list ligand GSH-Ag in reference examples 1 of the present invention2After S quantum dot system is to PC-12 cell marking
Fluorescence imaging figure.
Specific embodiment
More detailed explanation will hereafter be made to technical solution of the present invention.It is understood, however, that in model of the present invention
In enclosing, above-mentioned each technical characteristic of the invention and it is ok between each technical characteristic specifically described in below (e.g. embodiment)
It is combined with each other, to form a new or preferred technical solution.Due to space limitations, I will not repeat them here.
Recent studies have indicated that the area near-infrared II fluorescence (NIR-II, 1000-1700nm) has less in living tissue
Tissue resorption and scattering and lower tissue autofluorescence characteristic, can greatly improve fluorescence imaging tissue penetration depths and
Spatial resolution.> the tissue penetration depths of 1.5cm, the temporal resolution of 30ms can have been realized in living imaging at present
With~25 μm of spatial resolution, there is order of magnitude promotion compared with conventional fluorescent (400-900nm) imaging technique.In addition quantum dot also has
There is the optical property of voltage-sensitive, the red shift including quantum dot fluorescence emission peak, the reduction of spectrum widening and fluorescent emission intensity
Deng.Meanwhile quantum dot has very high resistance to photobleaching, and there is big quantum to produce single photon and two-photon excitation
Rate and absorption cross-section.It is greatly excellent that these excellent properties all determine that near-infrared II area's fluorescence quantum has as membrane probe
Gesture and application value.
In view of deficiency in the prior art, inventor is studied for a long period of time and is largely practiced, and is able to propose of the invention
Technical solution mainly passes through the size of the control area near-infrared II fluorescence quantum and pair of the hydrophilic, hydrophobic ligand of quantum dot surface
Functional modification has the function that compatible with cell membrane biological, can successfully imbed in the phospholipid bilayer of cell membrane, real
Now to the label of cell membrane and fluorescence imaging.Simultaneously as the area near-infrared II fluorescence (NIR-II, 1000-1700nm) is in living body
Characteristic in tissue with less tissue resorption and scattering and lower tissue autofluorescence, so that the tagging system is in living body
There is bigger application potential and value on the label of cell.As follows will in conjunction with attached drawing to the technical solution, its implementation process and
Principle etc. is further explained.
The area a kind of near-infrared II fluorescence quantum cell membrane tagging system packet that the one aspect of the embodiment of the present invention provides
Include: the area near-infrared II fluorescence quantum, and modification are connected to the hydrophily on the area near-infrared II fluorescence quantum surface and match
Body and hydrophobic ligand.Near-infrared II area's fluorescence quantum makes it be easier to be embedded in after hydrophilic/hydrophobic pair is ligand modified
Cell membrane marker and imaging are carried out in cell membrane.
In the present invention, using the distinctive size advantage of the area near-infrared II fluorescence quantum and luminosity, by surface
Functionalization realizes that the membrane structure to various different cells carries out highly efficient labeling and long-time is clear to improve biocompatibility
The effect of imaging.
In some embodiments, the hydrophilic ligand includes glutathione, dihydrolipoic acid, thioacetic acid, sulfydryl third
Any one in acid etc. or two or more combinations, but not limited to this.
Further, the hydrophobic ligand includes lauryl mercaptan (DT) ligand, but not limited to this.
Further, the double ligands of the hydrophilic/hydrophobic on the area near-infrared II fluorescence quantum surface can for glutathione/
Lauryl mercaptan ligand, dihydrolipoic acid/lauryl mercaptan ligand, thioacetic acid/lauryl mercaptan ligand or mercaptopropionic acid/lauryl mercaptan
Any one in ligand etc. or two or more combinations, but not limited to this.
In some embodiments, the launch wavelength of the area the near-infrared II (NIR-II, 1000-1700nm) fluorescence quantum
For the area near-infrared II, wave-length coverage is 1000~1700nm.Such as can be 1000nm, 1050nm, 1100nm, 1200nm,
1300nm, 1400nm, 1500nm, 1600nm or 1700nm.
Further, the area near-infrared II fluorescence quantum can be Ag2Se、Ag2S、InAs、InSb、GbSb、Ag2Te
With any one or the two or more combinations in PbS etc., but not limited to this.
Further, the partial size of the area near-infrared II fluorescence quantum is 1~10nm, preferably 2~5nm.
In the present invention, the area near-infrared II fluorescence quantum is after the double ligand surface functional modifications of hydrophilic/hydrophobic thin
Good biocompatibility and lower bio-toxicity are shown in born of the same parents' labeling process, while cell membrane is inserted into embedded mode
In phospholipid bilayer, reduce the influence and damage to cell membrane function, and played to the greatest extent to cell membrane mark
The effect of note and imaging.
The other side of the embodiment of the present invention additionally provides fluorescence quantum cell membrane marker system, the area aforementioned near-infrared II
The preparation method of system comprising: it modifies hydrophobic ligand, hydrophilic ligand respectively and is connected to the area near-infrared II fluorescence quantum
Surface obtains the area near-infrared II fluorescence quantum cell membrane tagging system.
In some embodiments, the preparation method specifically includes:
So that hydrophobic ligand modification is connected to the area near-infrared II fluorescence quantum surface, forms hydrophobic ligand quantum dot；
Hydrophilic ligand, hydrophobic ligand quantum dot and solvent are uniformly mixed, stirred, is obtained after centrifugation described close red
The outer area II fluorescence quantum cell membrane tagging system.
Among some more specifically case study on implementation, the step of preparation method mainly includes, is as follows:
So that hydrophobic ligand modification is connected to the area near-infrared II fluorescence quantum surface, forms hydrophobic ligand quantum dot
(also referred to as oily phase quantum dot)；
Oily phase quantum dot is done into surface hydrophilicity functionalized modification, metal Ion-hydrophilic Ligand modification is preferably glutathione, by gluathione
Peptide is added in chloroform-aqueous solution containing appropriate oily phase quantum dot, and it is thin to obtain the area near-infrared II fluorescence quantum for stirring after centrifugation
After birth tagging system (the also referred to as double ligand modified aqueous phase quantum points of hydrophilic/hydrophobic).
Further, the molar concentration rate of the hydrophilic ligand and hydrophobic ligand quantum dot is 200~1000:1, excellent
Be selected as 300~700:1, for example, can be 200:1,300:1,400:1,500:1,600:1,700:1,800:1,900:1 or
1000:1 etc., but not limited to this.
Further, the solvent includes chloroform-aqueous solution, and the volume ratio of the chloroform-Solution for Chloroform and water is 1
~10:1, preferably 3~7:1, such as can be 1:1,2:1,3:1,4:1,5:1,6:1,7:1,8:1,9:1 or 10:1 etc., but
It is without being limited thereto.
Further, the time of the stirring be 1~6h, preferably 2~5h, such as can be 1h, 2h, 3h, 4h, 5h or
6h etc., but not limited to this.
Further, the centrifugal force used that is centrifuged is 5000~15000g, preferably 8000~12000g, such as can
To be 8000g, 9000g, 10000g, 11000g or 12000g etc., but not limited to this.
The other side of the embodiment of the present invention additionally provides fluorescence quantum cell membrane marker system, the area aforementioned near-infrared II
Purposes of the system in cell membrane marker or fluorescence imaging field.
The other side of the embodiment of the present invention additionally provides a kind of cell membrane labeling method comprising: make difference wait mark
The cell of note is mixed with the area the aforementioned near-infrared II fluorescence quantum cell membrane tagging system of debita spissitudo, forms label system,
Total incubation is carried out later, makes the phospholipid bilayer of the area near-infrared II fluorescence quantum cell membrane tagging system insertion cell membrane
In layer, the label to cell membrane is realized.
The area near-infrared II fluorescence quantum is surface modified in an exemplary embodiments of the invention as shown in figure 1 and carefully
The schematic diagram of born of the same parents' label, wherein hydrophilic ligand selects glutathione, shows the parent with good biocompatibility in Fig. 1
The phosphatide that the water/hydrophobic double ligand modified area near-infrared II fluorescence quantum cell membrane tagging systems are successfully embedded in cell membrane is double to be divided
In sublayer, the process of cell membrane marker is realized.
Further, in the label system area near-infrared II fluorescence quantum cell membrane tagging system concentration be 1~
100μg·ml-1, preferably 10~50 μ gml-1, such as can be 1,5,10,15,20,50 or 100 μ gmL-1Deng, but not
It is limited to this.
Further, the cell can be PC-12 cell, spongiocyte or primary neuronal cell etc., but unlimited
In this.
Further, the condition of the label includes: that the label system is placed in the physiology salt that pH value is 6.8~7.5
In water, at 37 DEG C, 5%CO210~60min of co-incubation under concentration.
To sum up, the area near-infrared II fluorescence quantum cell membrane tagging system of the invention can overcome existing conventional fluorescent to visit
The defects of needle is easily quenched, and penetration depth is shallow and tissue autofluorescence is high, preferably can carry out the label of cell membrane and function is ground
Study carefully, the label especially on active somatic cell has huge application potential and value.
The technical solution and its effect taken for the present invention is further explained, with reference to embodiments with attached drawing to this hair
It is bright to be further described.It is understood that the specific embodiments described herein are only used for helping explain this hair
It is bright, it is not construed as to specific restriction of the invention.
In the examples where no specific technique or condition is specified, described technology or conditions according to the literature in the art,
Or carried out according to product description, agents useful for same or instrument are the conventional products that can be bought by regular distributor.
Embodiment 1
Hydrophily GSH-DT-Ag2The specific preparation method of S quantum dot:
The vial of a clean dried is taken, chloroform and deionized water are added thereto, according to certain molar concentration ratio
DT-Ag is added2S and glutathione (GSH), after magnetic agitation is mixed, continue to stir under the conditions of being protected from light, after take upper water
Mutually with the liquid at oil-water interfaces, upper strata aqueous phase is discarded through centrifugal treating, the primary centrifugation of deionization washing both obtains GSH- after being resuspended
DT-Ag2S water-soluble quantum dot.The GSH-DT-Ag prepared2Transmission electron microscope characterize data such as Fig. 2 institute of S hydrophily quantum dot
Show, average grain diameter is about 2nm.
Embodiment 2
Hydrophily DHLA-DT-Ag2The specific preparation method of S quantum dot:
The vial of a clean dried is taken, chloroform and deionized water are added thereto, according to certain molar concentration ratio
DT-Ag is added2S and dihydrolipoic acid (DHLA), after magnetic agitation is mixed, continue to stir under the conditions of being protected from light, after take
Liquid at layer water phase and oil-water interfaces discards upper strata aqueous phase through centrifugal treating, after the primary centrifugation resuspension of deionization washing both
DHLA-DT-Ag2S water-soluble quantum dot.Other hydrophilies parent/dredge the preparation method of double ligand quantum dots similarly.
Embodiment 3
The acquisition and culture of different cells involved in the present invention:
The present invention relates to the PC-12 cells used to be purchased from Chinese Academy of Sciences Shanghai cell bank, condition of culture are as follows: RPMI-1640
(GIBCO, article No. 31800022 add sodium bicarbonate 1.5g/L, glucose 2.5g/L, Sodium Pyruvate 0.11g/L), 85%；Heat
Inactivation horse serum, 10%；High-quality fetal calf serum, 5%.Gas phase: air, 95%；Carbon dioxide, 5%.Temperature: 37 DEG C.
The extraction step of primary neuron is as follows:
(1) newborn SD rat in birth for 24 hours impregnates 3-5min with 75% alcohol, and the HBSS for taking out pre-cooling cleans two
It is secondary, completely remove remaining alcohol；
(2) complete rat brain is taken out using tweezers, the DMEM-F12 of transposition pre-cooling (contains 10% horse serum, 1% blueness-
Streptomysin) in culture solution, the meninx and blood vessel of tissue are carefully removed with fine microforceps on ice, takes out complete hippocampus district's groups
It knits and is cut into cubed pieces, be placed in the DMEM-F12 culture solution of serum-free, separately add about 200 μ L TrypLE Express thereto
Digestive juice and 20 μ L DNase I digest 15min, every 5min rocks once in 6cm culture dish at 37 DEG C；
(3) 10% horse serum is added after 15min and terminates digestion, inclination culture dish stands 2-5min, removes supernatant, is added
2mL contains the DMEM-F12 of 10% calf serum, and slowly piping and druming 10 times, stand 2-5min on ice.Supernatant is taken, through 400 mesh cells
The screen to filtrate collects filtrate in 15mL centrifuge tube；
(4) the piping and druming step 2 time in step (3) is repeated, the cell suspension 1500rpm of collection is centrifuged 5min, in removal
Clearly, it is resuspended with the DMEM-F12 culture medium containing 10% calf serum, by 104Cell/ware density is inoculated into laser co-focusing ware
It is interior.
(5) 37 DEG C, 5% carbon dioxide incubator interior culture 4-6 hours shakes gently removal supernatant, with serum-free DMEM-
F12 culture medium cleans one time, and about 1mL Neurobasal Serum-free complete medium is added and is cultivated, partly changes liquid one within 2-3 days
It is secondary.
(6) after cultivating 84 hours, neuron can carry out cell membrane dyeing and fluorescence imaging.
For the quantum dot-labeled condition and method of different cells in the present embodiment:
To 3 kinds of cited cells in the present embodiment: PC-12 cell, spongiocyte and primary mouse neuronal cell are taken
Same flag condition.PH value be 7.4 physiological saline in 15 μ gmL-1Quantum dot jointly in 37 DEG C, 5%CO2It is dense
Degree is lower to cultivate 30min.Then 2 times are respectively washed with physiological saline and remove the extra quantum dot being not bound with.
Fig. 3 a- Fig. 3 c is label effect picture of the quantum dot system to above-mentioned 3 kinds different cells, witness marking in figure respectively
Uniform in effect can be clearly seen that membrane structure and complete cellular morphology.
Reference examples 1
In order to illustrate the double ligand modified quantum dots of hydrophilic/hydrophobic of the invention to the advantage of cell membrane marker, with reference to reality
The cell membrane flag condition in example 3 is applied, the quantum dot modified using only hydrophilic ligand, such as GSH-Ag2S quantum dot body
System, is marked PC-12 cell.Fig. 4 is GSH-Ag2S quantum dot system, can in Fig. 4 to the label effect picture of PC-12 cell
See that quantum dot is easy to reunite at one, uniformity is poor, can not clearly indicate the structure of cell membrane, and because quantum dot in
Intracellular a large amount of reunions impact cell function, show biggish cytotoxicity and are unfavorable for cell function and grind
Study carefully.The double ligand modified quantum dot systems of hydrophilic/hydrophobic in the present invention just successfully avoid quantum dot and are entering cell
After film it is a large amount of reunion and adversely affected caused by cell function, gather around the system in cell membrane marker and functional study field
There is bigger application value.
In conclusion the present invention is by above-mentioned technical proposal, the hydrophilic/hydrophobic that the present invention uses is double ligand modified close red
The outer area II fluorescence quantum cell membrane tagging system good biocompatibility, can successfully imbed the phospholipid bilayer of cell membrane
Interior, toxicity is low, has little effect to eucaryotic cell structure and function, and can to greatest extent to cell membrane be marked with it is glimmering
Light imaging.
In addition, inventor also utilizes the alternate embodiments such as listed other raw materials and other process conditions above
Various raw materials and corresponding process conditions in 1-2 have carried out corresponding test, the area obtained near-infrared II fluorescence quantum cell membrane mark
The performance of note system etc. is also ideal, substantially similar to Example 3.
It should be appreciated that the technical concepts and features of above-described embodiment only to illustrate the invention, its object is to allow be familiar with this
The personage of item technology cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all
Equivalent change or modification made by Spirit Essence according to the present invention, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of area near-infrared II fluorescence quantum cell membrane tagging system, characterized by comprising: the area near-infrared II fluorescence volume
It is sub-, and modify the hydrophilic ligand and hydrophobic ligand for being connected to the area near-infrared II fluorescence quantum surface.
2. the area near-infrared II according to claim 1 fluorescence quantum cell membrane tagging system, it is characterised in that: the parent
Aqueous ligand includes glutathione, dihydrolipoic acid, thioacetic acid, any one or two or more groups in mercaptopropionic acid
It closes；And/or the hydrophobic ligand includes lauryl mercaptan ligand.
3. the area near-infrared II according to claim 1 fluorescence quantum cell membrane tagging system, it is characterised in that: described close
The launch wavelength of the infrared area II fluorescence quantum is 1000~1700nm；Preferably, the area near-infrared II fluorescence quantum packet
Include Ag2Se、Ag2S、InAs、InSb、GbSb、Ag2Any one in Te and PbS or two or more combinations；
And/or the partial size of the area near-infrared II fluorescence quantum is 1~10nm, preferably 2~5nm.
4. the preparation method of the area near-infrared II fluorescence quantum cell membrane tagging system described in any one of claim 1-3,
It is characterized in that including: to modify hydrophobic ligand, hydrophilic ligand respectively to be connected to the area near-infrared II fluorescence quantum surface, obtains
Obtain the area near-infrared II fluorescence quantum cell membrane tagging system.
5. the preparation method according to claim 4, it is characterised in that specifically include:
So that hydrophobic ligand modification is connected to the area near-infrared II fluorescence quantum surface, forms hydrophobic ligand quantum dot；
Hydrophilic ligand, hydrophobic ligand quantum dot and solvent are uniformly mixed, stirred, the near-infrared II is obtained after centrifugation
Area's fluorescence quantum cell membrane tagging system.
6. preparation method according to claim 5, it is characterised in that: the hydrophilic ligand and hydrophobic ligand quantum dot
Molar ratio be 200~1000:1, preferably 300~700:1.
7. preparation method according to claim 5, it is characterised in that: the solvent includes chloroform-aqueous solution；Preferably,
The volume ratio of the chloroform-Solution for Chloroform and water is 1~10:1, preferably 3~7:1；
And/or the time of the stirring is 1~6h, preferably 2~5h；
And/or the centrifugal force that uses of being centrifuged is 5000~15000g, preferably 8000~12000g.
8. the area near-infrared II fluorescence quantum cell membrane tagging system described in any one of claim 1-3 in cell membrane marker or
Purposes in fluorescence imaging field.
9. a kind of cell membrane labeling method, characterized by comprising: make any one of cell and claim 1-3 to be marked institute
The mixing of the area near-infrared II fluorescence quantum cell membrane tagging system is stated, label system is formed, carries out total incubation later, is made described close
In the phospholipid bilayer of the infrared area II fluorescence quantum cell membrane tagging system insertion cell membrane, the mark to cell membrane is realized
Note.
10. cell membrane labeling method according to claim 9, it is characterised in that: the area near-infrared II in the label system
The concentration of fluorescence quantum cell membrane tagging system is 1~100 μ gml-1, preferably 10~50 μ gml-1；
And/or the condition of the label includes: to be placed in the label system in the physiological saline that pH value is 6.8~7.5,
37 DEG C, 5%CO210~60min of co-incubation under concentration.
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| CN113004889A (en) * | 2021-03-09 | 2021-06-22 | 大连民族大学 | Method for synthesizing near-infrared fluorescence/photothermal/photoacoustic silver sulfide quantum dots |
| CN113981004A (en) * | 2021-11-01 | 2022-01-28 | 中国科学院苏州纳米技术与纳米仿生研究所 | Genetic coding nano probe for cell membrane potential detection and preparation method and application thereof |
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| CN102277157A (en) * | 2011-05-30 | 2011-12-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Near-infrared silver sulphide quantum dot as well as preparation method and application thereof |
| CN104288787A (en) * | 2014-09-22 | 2015-01-21 | 东南大学 | Washing-free long-time cell membrane fluorescence imaging reagent and preparation method thereof |
| CN106753344A (en) * | 2016-11-25 | 2017-05-31 | 清华大学 | Silver sulfide quantum dot and preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102277157A (en) * | 2011-05-30 | 2011-12-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Near-infrared silver sulphide quantum dot as well as preparation method and application thereof |
| CN104288787A (en) * | 2014-09-22 | 2015-01-21 | 东南大学 | Washing-free long-time cell membrane fluorescence imaging reagent and preparation method thereof |
| CN106753344A (en) * | 2016-11-25 | 2017-05-31 | 清华大学 | Silver sulfide quantum dot and preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113004889A (en) * | 2021-03-09 | 2021-06-22 | 大连民族大学 | Method for synthesizing near-infrared fluorescence/photothermal/photoacoustic silver sulfide quantum dots |
| CN113981004A (en) * | 2021-11-01 | 2022-01-28 | 中国科学院苏州纳米技术与纳米仿生研究所 | Genetic coding nano probe for cell membrane potential detection and preparation method and application thereof |
| CN113981004B (en) * | 2021-11-01 | 2024-02-09 | 中国科学院苏州纳米技术与纳米仿生研究所 | Genetically encoded nano probe for cell membrane potential detection and preparation method and application thereof |
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