CN113698935B - CdZnSe/Mn ZnS QDs, synthetic method and application thereof - Google Patents
CdZnSe/Mn ZnS QDs, synthetic method and application thereof Download PDFInfo
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Abstract
The synthesis method of the CdZnSe/Mn: znS QDs provided by the application takes the CdZnSe quantum dot prepared by a water bath method as a core, and then Mn is carried out 2+ Finally, wrapping ZnS shell layers on the surfaces of the CdZnSe quantum dots through Mn 2+ The method of doping in the core can well ensure Mn 2+ The emitted light-emitting efficiency is doped, so that a bicolor quantum dot with high light-emitting efficiency is obtained, and lattice mismatch among ZnSe, znS and MnS can be reduced due to low difference of lattice parameters, so that quantum yield and stability of the quantum dot are improved. The synthesized CdZnSe/Mn: znS QDs can be directly used as a ratio fluorescent probe for pH monitoring in living cells.
Description
Technical Field
The application relates to the field of nanomaterials and biomedical analytical chemistry, in particular to a synthesis method of CdZnSe/Mn: znS QDs, the CdZnSe/Mn: znS QDs and application thereof.
Background
Conventional nano-fluorescent probes rely mainly on the intensity variation of the individual fluorescence and are susceptible to numerous factors such as instrument fluctuations, probe concentration fluctuations, and non-specific binding. These factors can lead to strong non-specific background signals and high false positives (x.deng et al, anal. Chem.2020,92,15908.). To solve this problem, researchers have proposed a ratio fluorescent probe method based on two different fluorescent peak intensity ratios, one of which is identified as a signal and the other as a reference. The method can improve the signal-to-noise ratio so as to improve the detection accuracy. Furthermore, the method of ratiometric fluorescence allows for simple, rapid visual analysis of analytes through color changes. Quantum dots have excellent optical and chemical properties such as high luminous efficiency, narrow and symmetrical emission peak, wide absorption peak, large stokes shift and strong resistance to photobleaching, and thus have been widely used in biosensing (i.l. medintz et al, nat. Mater 2005,4,435). However, most ratiometric fluorescent probes are constructed by covalent coupling reactions of quantum dots with other fluorophores (small organic molecules, fluorescent proteins, or other nanomaterials, etc.), a process that reduces the luminous efficiency and stability of the quantum dots. In earlier work, we developed a ratiometric fluorescent probe based on DNA functional quantum dots (g.mao et al, anal. Chem.2017,89,11628.). The method has mild conditions, and can effectively protect the quantum yield and stability of the quantum dots. However, this method is expensive and the stokes shift of the dye molecule is small. Therefore, it is of great importance to construct a simple and efficient method for preparing a ratio-type fluorescent probe.
Mn due to its own but particle dual fluorescence emission characteristics 2+ Doped quantum dots have received widespread attention in recent years. Mn in common use 2+ The preparation method of the doped quantum dot comprises the steps of preparing Mn 2+ Rational doping into wide bandgap host materials ZnSe or ZnS, the dual fluorescence emission of which comprises a high energy bandgap host emission and a low energy Mn 2+ Doped emission, respectively due to exciton emission and Mn of the host 2 + Doped with 4 T 1 → 6 A 1 Internal d-d transition emission. Lu et al developed a method for preparing double emission ZnS: mn 2+ QD methods and explored their use in biological analysis by means of energy transfer, chemical reactions and electron transfer (l.lu et al, anal. Chem.2014,86,6188.). There have been many researchers developing a number of methods for preparing dual fluorescence emission Mn 2+ Method of doping perovskite materials, and can be modifiedModulating excitons and Mn by varying laser excitation repetition rate or pulse intensity, etc 2+ The relative emission intensity between the dopants (q.sun et al, j.am. Chem. Soc.2019,141, 20089.). However, preparing water-soluble bifluorescent emissive quantum dots with high quantum yield and good stability remains somewhat challenging, probably due to Mn 2+ The fluorescence of the doping is very dependent on Mn 2+ Concentration and Mn in bulk Quantum dot 2+ Is a spatial location of (c).
Intracellular pH plays a critical role in regulating the function and activity of cells and organisms, including signal transduction, proliferation and apoptosis (A. Steineger et al, chem. Rev.2020,120, 12357.). Intracellular pH abnormalities can lead to significant changes in metabolism, and studies have shown a link between the two. In addition, the pH value is also closely related to diseases such as cancers (tumorigenesis and metastasis) and Alzheimer's disease (C.Ding et al, anal. Chem.2019,91,7181.). Therefore, the method has great significance for accurately monitoring the fluctuation of the pH value in the living cells. To date, a variety of techniques have been used to monitor pH changes, including electrochemical, nuclear magnetic resonance, and fluorescence (t.sakata et al, anal.Chem.2018,90,12731;H.Chen et al,Anal.Chem.2018,90,7056;X.Tang et al,Anal.Chem.2020,92,16293.). Among the various techniques described above, fluorescence shows unique advantages, including good signal background, and the ability of real-time analysis of individual living cells.
Disclosure of Invention
In view of this, it is necessary to provide a method for synthesizing CdZnSe/Mn: znS QDs having single-particle dual-fluorescence emission properties and used for the imaging analysis of the pH in HeLa cells.
In order to solve the problems, the application adopts the following technical scheme:
the application provides a method for synthesizing CdZnSe/Mn: znS QDs, which comprises the following steps:
preparing CdZnSe QDs;
and mixing the CdZnSe QDs, manganese acetate, a mixed solution B and a sodium sulfide solution, and then heating in a water bath to react to obtain the CdZnSe/Mn: znS QDs, wherein the mixed solution B is obtained by dissolving zinc chloride and glutathione in deionized water.
In some of these embodiments, the step of preparing CdZnSe QDs specifically comprises:
dissolving cadmium chloride and glutathione in deionized water to obtain a mixed solution A;
dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B;
adjusting the pH value of the mixed solution A and the mixed solution B to 9.0-11.0;
centrifuging selenium powder and sodium borohydride at room temperature until the solution is colorless, so as to obtain a mixed solution C;
mixing the mixed solution C, the mixed solution A and the mixed solution B, and then heating in a water bath to react to obtain a CdZnSe QDs solution;
and separating and purifying the CdZnSe QDs solution to obtain the CdZnSe QDs.
In some embodiments, in the step of dissolving cadmium chloride and glutathione in deionized water to obtain the mixed solution A, the molar ratio of the cadmium chloride to the glutathione is 1 (0.5-1).
In some embodiments, in the step of dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B, the molar ratio of the zinc chloride to the glutathione is 1 (0.5-2).
In some embodiments, the molar ratio of the mixed solution A to the mixed solution B is 1 (0.1-0.2).
In some embodiments, in the step of adjusting the pH value of the mixed solution a and the mixed solution B to 9.0-11.0, specifically: and regulating the pH value of the mixed solution A and the mixed solution B to 9.0-11.0 by using sodium hydroxide solution, wherein the concentration of the sodium hydroxide solution is 0.5-5 mol/L.
In some embodiments, in the step of centrifugally reacting selenium powder and sodium borohydride at room temperature until the solution is colorless to obtain a mixed solution C, the molar ratio of the selenium powder to the sodium borohydride is 1:1-4.
In some embodiments, in the step of mixing the mixed solution C, the mixed solution a and the mixed solution B and then heating in a water bath to react, the CdZnSe QDs solution is specifically: and (3) uniformly mixing and stirring the mixed solution C, the mixed solution A and the mixed solution B, transferring the mixture into a water bath kettle, and reacting at 95 ℃ for 30min to obtain the CdZnSe QDs.
In some of these embodiments, cdCl is present in the mixed solution of the mixed solution C, the mixed solution a, and the mixed solution B 2 The concentration is 0.001-0.0625 mM.
In some embodiments, in the step of separating and purifying the CdZnSe QDs solution to obtain CdZnSe QDs, specifically:
and (3) carrying out centrifugal purification on the CdZnSe QDs solution by using an ultrafiltration tube, removing waste liquid, adding ultrapure water, washing and centrifuging, inversely centrifuging the ultrafiltration tube to obtain CdZnSe QDs, and storing at 4 ℃ for standby, wherein the molecular retention of the ultrafiltration tube is 30KD, the washing and centrifuging times are 4 times, the rotating speed during each centrifuging is 10000r/min, and the centrifuging time is 10min.
In some embodiments, in the step of mixing the CdZnSe QDs, manganese acetate, the mixed solution B and sodium sulfide solution, and then heating in a water bath to react, the CdZnSe/Mn: znS QDs is specifically:
sequentially adding manganese acetate, the mixed solution B and the sodium sulfide solution into the CdZnSe QDs, uniformly mixing, transferring to a water bath kettle, and reacting at 95 ℃ for 30min to perform water bath heating reaction to obtain the CdZnSe/Mn ZnS QDs.
In addition, the application also provides a CdZnSe/Mn: znS QDs synthesized by the synthesis method of the CdZnSe/Mn: znS QDs.
In addition, the application also provides an application of the CdZnSe/Mn: znS QDs in live cell pH imaging.
By adopting the technical scheme, the application has the following technical effects:
the synthesis method of the CdZnSe/Mn: znS QDs provided by the application takes the CdZnSe QDs quantum dots prepared by a water bath method as cores, and then Mn is carried out 2+ Finally, wrapping ZnS shell layers on the surfaces of the CdZnSe quantum dots through Mn 2+ The method of doping in the core can well ensure Mn 2+ The emitted light-emitting efficiency is doped, so that a bicolor quantum dot with high light-emitting efficiency is obtained, and lattice mismatch among ZnSe, znS and MnS can be reduced due to low difference of lattice parameters, so that quantum yield and stability of the quantum dot are improved.
The synthesized CdZnSe/Mn: znS QDs can be directly used as a ratio fluorescent probe for pH monitoring in living cells, when the pH value of HeLa cells is changed from 9.0 to 5.0, the color in the living cells is gradually changed from red to pink, and finally, the color is blue, so that the CdZnSe/Mn: znS QDs has good effect on monitoring the pH in the living cells, and can provide a certain material foundation and analysis platform for other biosensing and biological imaging analysis.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following description will briefly explain the embodiments of the present application or the drawings used in the description of the prior art, and it is obvious that the drawings described below are only some embodiments of the present application, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a step flow chart of a method for synthesizing CdZnSe/Mn: znS QDs according to an embodiment of the application.
FIG. 2 is a schematic diagram of the synthesis of CdZnSe/Mn: znS QDs and their use in live intracellular pH imaging, provided by the application.
FIG. 3 is a fluorescence spectrum of CdZnSe/Mn: znS QDs synthesized in example 2 of the present application.
FIG. 4 is a graph showing the results of ultraviolet-visible spectrum characterization of the CdZnSe QDs prepared in example 1 and the CdZnSe/Mn: znS QDs prepared in example 2 of the present application.
FIG. 5 is a graph showing the fluorescence lifetime of CdZnSe/Mn: znS QDs prepared in example 2 of the present application.
FIG. 6 is a graph showing the characterization results of various materials of CdZnSe/Mn: znS QDs prepared in example 2 of the present application.
FIG. 7 is a schematic diagram showing the examination of the stability of CdZnSe/Mn: znS QDs synthesized in example 2 of the present application.
FIG. 8 is a cytotoxicity examination of CdTe QDs, cdZnSe QDs and CdZnSe/Mn: znS QDs prepared in example 2 of the present application.
FIG. 9 shows the use of CdZnSe/Mn: znS QDs prepared in example 2 of the present application for pH measurement.
FIG. 10 is a schematic diagram showing the optimization of the quantum dot usage amount when the CdZnSe/Mn: znS QDs synthesized in example 2 of the present application are used for HeLa cell imaging.
FIG. 11 is a graph showing the results of pH imaging of CdZnSe/Mn: znS QDs synthesized in example 2 for HeLa cells according to the present application.
Detailed Description
Embodiments of the present application are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present application and should not be construed as limiting the application.
In the description of the present application, it should be understood that the directions or positional relationships indicated by the terms "upper", "lower", "horizontal", "inner", "outer", etc., are based on the directions or positional relationships shown in the drawings, are merely for convenience in describing the present application and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present application.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present application, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
The present application will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present application more apparent.
Referring to FIG. 1, a step flow chart of a method for synthesizing CdZnSe/Mn: znS QDs according to an embodiment of the application includes the following steps:
step S110: preparing CdZnSe QDs;
in some of these embodiments, the preparation of CdZnSe QDs by a water bath process specifically comprises:
step S111: and (3) dissolving cadmium chloride and glutathione in deionized water to obtain a mixed solution A.
In some of these embodiments, in the step of dissolving cadmium chloride and glutathione in deionized water to obtain mixed solution A (Cd-GSH), the molar ratio of the cadmium chloride to the glutathione is 1 (0.5-1).
Step S112: and dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B.
In some of these embodiments, in the step of dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B (Zn-GSH), the molar ratio of the zinc chloride to the glutathione is 1 (0.5-2).
Step S113: and regulating the pH value of the mixed solution A and the mixed solution B to 9.0-11.0.
In some embodiments, the molar ratio of the mixed solution A to the mixed solution B is 1 (0.1-0.2).
Further, in the step of adjusting the pH value of the mixed solution a and the mixed solution B to 9.0 to 11.0, specifically: and regulating the pH value of the mixed solution A and the mixed solution B to 9.0-11.0 by using sodium hydroxide solution, wherein the concentration of the sodium hydroxide solution is 0.5-5 mol/L.
Step S114: and (3) carrying out centrifugal reaction on the selenium powder and sodium borohydride at room temperature until the solution is colorless, so as to obtain a mixed solution C.
In some embodiments, in the step of centrifugally reacting selenium powder and sodium borohydride at room temperature until the solution is colorless to obtain a mixed solution C, the molar ratio of the selenium powder to the sodium borohydride is 1:1-4.
Step S115: and mixing the mixed solution C, the mixed solution A and the mixed solution B, and then heating in a water bath to react to obtain the CdZnSe QDs solution.
In some embodiments, in the step of mixing the mixed solution C, the mixed solution a and the mixed solution B and then heating in a water bath to react, the CdZnSe QDs solution is specifically: and (3) uniformly mixing and stirring the mixed solution C, the mixed solution A and the mixed solution B, transferring the mixture into a water bath kettle, and reacting at 95 ℃ for 30min to obtain the CdZnSe QDs.
Further, cdCl is contained in the mixed solution C, the mixed solution a, and the mixed solution B 2 The concentration is 0.001-0.0625 mM.
Step S116: and separating and purifying the CdZnSe QDs solution to obtain the CdZnSe QDs.
In some embodiments, in the step of separating and purifying the CdZnSe QDs solution to obtain CdZnSe QDs, specifically:
and (3) carrying out centrifugal purification on the CdZnSe QDs solution by using an ultrafiltration tube, removing waste liquid, adding ultrapure water, washing and centrifuging, inversely centrifuging the ultrafiltration tube to obtain CdZnSe QDs, and storing at 4 ℃ for standby, wherein the molecular retention of the ultrafiltration tube is 30KD, the washing and centrifuging times are 4 times, the rotating speed during each centrifuging is 10000r/min, and the centrifuging time is 10min.
It can be understood that the synthesis of the CdZnSe QDs in the embodiment adopts a water bath method, so that the energy consumption is low, the efficiency is high, the performance of the quantum dots is good, the emission spectrum is easy to regulate and control, and the synthesis method is simple, the synthesis process of the quantum dots is greatly simplified, and the cost is saved.
Step S120: and mixing the CdZnSe QDs, manganese acetate, a mixed solution B and a sodium sulfide solution, and then heating in a water bath to react to obtain the CdZnSe/Mn: znS QDs, wherein the mixed solution B is obtained by dissolving zinc chloride and glutathione in deionized water.
In some embodiments, in the step of mixing the CdZnSe QDs, manganese acetate, the mixed solution B and sodium sulfide solution, and then heating in a water bath to react, the CdZnSe/Mn: znS QDs is specifically:
sequentially adding manganese acetate, the mixed solution B and the sodium sulfide solution into the CdZnSe QDs, uniformly mixing, transferring to a water bath kettle, and reacting at 95 ℃ for 30min to perform water bath heating reaction to obtain the CdZnSe/Mn ZnS QDs.
It can be understood that the synthesis of the CdZnSe/Mn: znS QDs provided by the application adopts a water bath method, and has low energy consumption and high efficiency. In addition, by Mn 2+ The method of doping in the core can well ensure Mn 2+ The light-emitting efficiency of the doped emission is improved, so that the bicolor quantum dot with high light-emitting efficiency is obtained.
In addition, in the synthesis process of the CdZnSe/Mn: znS QDs, the method of wrapping with ZnS layers can reduce Mn 2+ The spontaneous quenching of doped fluorescence can improve the optical and chemical stability of CdZnSe/Mn: znS QDs and promote the application of the CdZnSe/Mn in life analysis.
The synthesized CdZnSe/Mn: znS QDs can be directly used as a ratio fluorescent probe for pH monitoring in living cells, when the pH value of HeLa cells is changed from 9.0 to 5.0, the color in the living cells is gradually changed from red to pink, and finally, the color is blue, so that the CdZnSe/Mn: znS QDs has good effect on monitoring the pH in the living cells, and can provide a certain material foundation and analysis platform for other biosensing and biological imaging analysis.
Referring to FIG. 2, a schematic diagram of the synthesis of CdZnSe/Mn: znS QDs and their use in live-cell pH imaging is provided.
The synthesis method of CdZnSe/Mn: znS QDs provided by the application firstly adopts a water bath method to prepare the CdZnSe QDs, and then Mn is adopted to synthesize the CdZnSe/Mn: znS QDs 2+ The single-particle double-fluorescence-emission CdZnSe/Mn: znS QDs is obtained by doping the single-particle double-fluorescence-emission CdZnSe/Mn: znS QDs on the surface of the CdZnSe QDs and finally wrapping a layer of ZnS on the surface of the CdZnSe QDs, wherein the CdZnSe/Mn: znS QDs can be aggregated under an acidic condition, and the fluorescence emitted by the main body of the CdZnSe QDs has obvious aggregation-induced quenching phenomenon, and Mn 2+ The doped fluorescence is hardly affected after aggregation, so that single-particle double-fluorescence-emitted CdZnSe/Mn: znS QDs have good response relation to pH, as shown in FIG. 2, cdZnSe/Mn: znS QDs appear red in acidic cells (as indicated in the figure) and alkaline cellsBluish violet (as indicated in the figure) to enable accurate monitoring of the pH in living cells.
The process of the present application is further described in connection with specific examples to provide a further understanding of the present application by those skilled in the art, but these examples are merely preferred embodiments and the scope of the claims is not limited to the examples.
Example 1
The embodiment provides a synthesis method of CdZnSe quantum dots, which comprises the following steps:
1) Dissolving 0.025mmol of cadmium chloride and 0.05mmol of glutathione in deionized water to obtain a mixed solution A, dissolving 0.025mmol of zinc chloride and 0.05mmol of glutathione in deionized water to obtain a mixed solution B, and regulating the pH values of the mixed solution A and the mixed solution B to 11.0 by using a sodium hydroxide solution; taking 0.01mmol of selenium powder and 0.02mmol of sodium borohydride, adding the selenium powder and the sodium borohydride into a 2mL centrifuge tube, and reacting for 10 minutes at room temperature to obtain a mixed solution C; mixing the solution A and the solution B according to a ratio of 1:9, rapidly transferring the mixed solution C into the mixed solution of the solution A and the solution B, uniformly stirring, transferring into a water bath kettle, and reacting for 30min at 95 ℃ to obtain the CdZnSe quantum dot;
2) And (3) carrying out centrifugal purification on the obtained CdZnSe quantum dot solution by using an ultrafiltration tube with a molecular retention of 30KD, pouring out waste liquid, adding ultrapure water for washing and centrifuging, circularly washing and centrifuging for 4 times, reversely centrifuging the ultrafiltration tube to obtain a CdZnSe quantum dot pure product, storing at 4 ℃ for standby, and diluting the CdZnSe quantum dot pure product into an aqueous solution with a required concentration when required.
Example 2
The embodiment provides a synthesis method of CdZnSe/Mn: znS quantum dots, which comprises the following steps:
taking purified CdZnSe QDs (2 mu M), adding MnCl with different concentrations 2 (0,0.01,0.02,0.04,0.1,0.2,0.4. Mu.M), then the mixed solution B of 1) is added, and finally Na is added 2 S solution. And rapidly and uniformly stirring the mixed solution, transferring the mixed solution into a water bath kettle, and reacting for 30min at 80 ℃ to obtain the CdZnSe: mn/ZnS QDs. Centrifugal ultrafiltration is carried out on the reaction solution by using a 50KD ultrafiltration tube, and the ultrafiltration is carried outThe filtration time was 10min at 8000rpm. And (5) placing the obtained CdZnSe/Mn: znS QDs in a refrigerator at the temperature of 4 ℃ for standby.
Example 3
The embodiment provides a performance characterization method of CdZnSe/Mn: znS QDs, which comprises the following steps:
the light stability was characterized by taking 100nM CdZnSe/Mn: znS QDs, irradiating under a 350V xenon lamp at a wavelength of 350nM for 2 hours, and continuously measuring the intensities of CdZnSe/Mn: znS QDs at 471 and 606 nM. As a control, 100nM CdTe QDs (610 nM) was used. CdZnSe/Mn ZnS QDs were stored in a refrigerator at 4℃and the fluorescence intensity was measured every 7 days to examine the stability. A20 nM CdZnSe/Mn: znS QDs was taken, and 0.2% and 1% bovine fetal serum was added, respectively, to examine the effect of serum on the CdZnSe/Mn: znS QDs, with 20nM CdZnSe QDs as a control.
Example 4
The embodiment provides a method for investigating the biocompatibility of CdZnSe/Mn: znS QDs, which comprises the following steps:
HeLa cells were incubated in medium containing 10% bovine fetal serum and 1% antibiotics. After the cells are full, they are hydrolyzed with trypsin and plated in 96-well plates at 37℃and 5% CO 2 Incubate under conditions for 24h. CdTe QDs, cdZnSe QDs and CdZnSe/Mn: znS QDs with different concentrations are respectively added into the 96-well plate, and incubation is continued for 24 hours. MTS reagent was added and finally absorbance at 490nm was measured with a microplate reader.
Example 5
The embodiment provides a method for detecting pH by using CdZnSe/Mn: znS QDs, which comprises the following steps:
preparing Tris buffer solutions with different pH values, adding 100nM CdZnSe/Mn: znS QDs into the Tris buffer solutions with different pH values, and respectively testing fluorescence spectra. Adding 100nM CdZnSe/Mn: znS QDs into Tris buffer with pH of 4.0, and testing fluorescence spectrum; the pH value is adjusted to 9.0 by NaOH, and the fluorescence spectrum is tested; the pH value is adjusted to 4.0 by NaOH, and the fluorescence spectrum is tested; the cycle was repeated 6 times, and finally, the change of the fluorescence intensity ratio was analyzed.
Example 6
The present example provides a method for imaging analysis of pH in HeLa cells using CdZnSe/Mn ZnS QDs, the imaging method comprising the steps of:
HeLa cells were incubated in medium containing 10% bovine fetal serum and 1% antibiotics. After incubation, the medium was gently blotted and the cells were rinsed twice with 1mL PBS. Then, heLa cells were incubated in a medium containing different concentrations of CdZnSe/Mn: znS QDs for 2h at 37℃to optimize the amount of CdZnSe/Mn: znS QDs. The CdZnSe/Mn ZnS QDs-containing medium was gently aspirated and the cells were rinsed three times with 1mL of medium. HeLa cells were observed with confocal microscopy using fresh medium instead of old medium.
HeLa cells were incubated in medium containing 10% bovine fetal serum and 1% antibiotics. After incubation, the medium was gently blotted and the cells were rinsed twice with 1mL PBS. Then, heLa cells were further incubated in a medium containing different concentrations of CdZnSe/Mn: znS QDs for 2 hours at 37 ℃. The CdZnSe/Mn ZnS QDs-containing medium was gently aspirated and the cells were rinsed three times with 1mL of medium. HeLa cells were observed with confocal microscopy after 30 minutes incubation with buffer solutions containing Nigericin at different pH values instead of old medium.
FIG. 3 is a graph showing the fluorescence spectrum of CdZnSe/Mn: znS QDs synthesized in example 2. When doped with Mn 2+ The concentration gradually increases, the fluorescence of CdZnSe QDs gradually decreases, and Mn 2+ The doping fluorescence of (2) is gradually enhanced; under the irradiation of an ultraviolet lamp, the color gradually changes from blue to purple and finally changes into pink. Thus, the quantum dot can pass through Mn 2+ The change of the concentration realizes the accurate regulation and control of the intensities of the two fluorescence peaks.
Referring to FIG. 4, graphs of the ultraviolet-visible spectrum characterization results of the CdZnSe QDs prepared in example 1 and the CdZnSe/Mn: znS QDs prepared in example 2 are shown. The main absorption peak of CdZnSe QDs is about 420nm, while the main absorption peak of CdZnSe/Mn: znS QDs is about 300 nm. The change in the position of the ultraviolet absorption peak initially demonstrates successful encapsulation of the ZnS layer.
Referring to FIG. 5, there is a graph showing the fluorescence lifetime of CdZnSe/Mn: znS QDs prepared in example 2. With Mn 2+ The fluorescence lifetime of the host CdZnSe host gradually decreases due to the increase of the doping amount, because of the CdZnSe and Mn 2+ Energy transfer occurs therebetween.
Referring to FIG. 6, a graph showing characterization results of various materials of CdZnSe/Mn: znS QDs prepared in example 2 is shown. A in FIG. 6 is a graph of the characterization result of a transmission electron microscope, which shows that CdZnSe/Mn: znS QDs have better dispersibility, uniform particle size and about 4.3nm. In FIG. 6, B is the hydrated particle size distribution of CdZnSe/Mn: znS QDs under different pH conditions, and the result shows that the hydrated particle size of CdZnSe/Mn: znS QDs is about 9.6nm at pH 9.0 and is as high as 5036nm at pH 4.0, indicating that the obvious aggregation phenomenon of CdZnSe/Mn: znS QDs occurs at pH 4.0. In FIG. 6, C is a graph of the surface potential results for CdZnSe QDs and CdZnSe/Mn: znS QDs, both quantum dots show negative charges, and the surface potentials are-21.3 and-16.4 mV, respectively. FIG. 6D is a graph showing the infrared absorption spectrum of CdZnSe QDs and CdZnSe/Mn: znS QDs at 2593cm compared with Glutathione (GSH) -1 The disappearance of the peak of (2) indicates that the thiol group in GSH is complexed with the metal ion. In FIG. 6, E is an X-ray crystal diffraction characterization result graph of CdZnSe QDs and CdZnSe/Mn: znS QDs, and the result shows that the CdZnSe QDs have two crystal forms of CdSe and ZnSe, and the CdZnSe/Mn: znS QDs have obvious red shift compared with the CdZnSe QDs, so that the successful wrapping of the ZnS layer is shown. FIG. 6 is a graph of the X-ray photoelectron spectroscopy results of CdZnSe QDs and CdZnSe/Mn: znS QDs showing Mn 2+ After doping of the ZnS layer and encapsulation of the Cd layer 2+ And Se (Se) 2- Is obviously reduced in content of S 2- And Zn 2+ Wherein Cd is significantly increased in concentration 2+ The reduced concentration may reduce the toxicity of the quantum dots.
Referring to FIG. 7, the stability of the CdZnSe/Mn: znS QDs synthesized in example 2 was examined, and the procedure was examined as in example 3. In FIG. 7, A is the light stability investigation of CdTe QDs and CdZnSe/Mn: znS QDs, and the result shows that the main fluorescence and doped fluorescence of CdZnSe/Mn: znS QDs are hardly reduced, and as a comparison, the fluorescence of CdTe QDs only has 60% after being irradiated for two hours, thus showing that the double-color fluorescence of CdZnSe/Mn: znS QDs has good optical stability. In FIG. 7, B is the time stability of CdZnSe/Mn: znS QDs, and the result shows that the host fluorescence and the doped fluorescence of CdZnSe/Mn: znS QDs hardly changed within 35 days of storage. In FIG. 7, C is the effect of bovine serum on CdZnSe QDs fluorescence, and the result shows that the effect of bovine serum on CdZnSe QDs fluorescence is obviously enhanced and may interfere with the practical application effect of CdZnSe QDs. In FIG. 7, D is the effect of bovine serum on CdZnSe/Mn: znS QDs fluorescence, and the result shows that 1% of bovine serum has no effect on the main fluorescence and doped fluorescence of CdZnSe/Mn: znS QDs, indicating that CdZnSe/Mn: znS QDs has better anti-environmental interference capability.
Referring to FIG. 8, cytotoxicity of CdTe QDs, cdZnSe QDs and CdZnSe/Mn: znS QDs prepared in example 2 was examined, and specific procedure is described in example 4. As shown in FIG. 8, at a concentration of 500nM, both CdZnSe QDs and CdZnSe/Mn: znS QDs were not significantly cytotoxic, whereas CdTe QDs as a control group had a cell viability of only 30%.
Referring to FIG. 9, for the determination of pH value using CdZnSe/Mn: znS QDs prepared in example 2, see example 5 for specific steps. FIG. 9A is a graph of fluorescence spectra of CdZnSe/Mn: znS QDs at different pH values, showing that the host fluorescence of CdZnSe/Mn: znS QDs is gradually increased while the Mn2+ doped fluorescence is substantially unchanged when the pH value is gradually increased. The inset of fig. 9 a shows the ratio of the intensities of the host fluorescence to mn2+ doped fluorescence at different pH conditions, indicating that the higher the pH, the greater the ratio of fluorescence intensities. In FIG. 9, B is the fluorescence intensity ratio of CdZnSe/Mn: znS QDs in the process of repeatedly changing the pH value from 4 to 9, and the result shows that the fluorescence intensity ratio of the CdZnSe/Mn: znS QDs solution is basically unchanged after a plurality of pH changes, so that the CdZnSe/Mn: znS QDs has good pH reciprocability and is suitable for the construction of a pH sensor.
Referring to FIG. 10, for the use of the synthesized CdZnSe/Mn ZnS QDs in HeLa cell imaging, the quantum dot is optimized, and the specific procedure is shown in example 6. The results show that the more quantum dots are added, the more obvious the fluorescence intensity in HeLa cells is; when the dosage of the quantum dots is 400nM, the effect is equivalent to that of the quantum dots with the dosage of 300nM, which indicates that the dosage of the quantum dots is basically saturated.
Referring to FIG. 11, a graph of the results of the synthesis of CdZnSe/Mn: znS QDs for HeLa cell pH imaging in example 2 is shown, for specific steps in example 6. As shown in the figure, when the intracellular pH is 5.0, the intracellular color is mainly red, when the pH is gradually marked as 9.0 from 5.0, the intracellular color is gradually marked as purple from red, and finally, the intracellular color is blue, which shows that CdZnSe/Mn: znS QDs can be well used for imaging analysis of the pH in living cells.
The synthesis of the CdZnSe/Mn: znS QDs provided by the embodiment of the application adopts a water bath method, and has low energy consumption and high efficiency. In addition, by Mn 2+ The method of doping in the core can well ensure Mn 2+ The light-emitting efficiency of the doped emission is improved, so that the bicolor quantum dot with high light-emitting efficiency is obtained.
In addition, in the synthesis process of the CdZnSe/Mn: znS QDs, the method of wrapping with ZnS layers can reduce Mn 2+ The spontaneous quenching of doped fluorescence can improve the optical and chemical stability of CdZnSe/Mn: znS QDs and promote the application of the CdZnSe/Mn in life analysis.
The synthesized CdZnSe/Mn: znS QDs can be directly used as a ratio fluorescent probe for pH monitoring in living cells, when the pH value of HeLa cells is changed from 9.0 to 5.0, the color in the living cells is gradually changed from red to pink, and finally, the color is blue, so that the CdZnSe/Mn: znS QDs has good effect on monitoring the pH in the living cells, and can provide a certain material foundation and analysis platform for other biosensing and biological imaging analysis.
The foregoing description of the preferred embodiments of the present application has been provided for the purpose of illustrating the general principles of the present application and is not to be construed as limiting the scope of the application in any way. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application, and other embodiments of the present application as will occur to those skilled in the art without the exercise of inventive faculty, are intended to be included within the scope of the present application.
Claims (13)
1. The synthesis method of the CdZnSe/Mn: znS QDs is characterized by comprising the following steps of:
preparing CdZnSe QDs;
and mixing the CdZnSe QDs, manganese chloride, a mixed solution B and a sodium sulfide solution, and then heating in a water bath to react to obtain the CdZnSe/Mn: znS QDs, wherein the mixed solution B is obtained by dissolving zinc chloride and glutathione in deionized water.
2. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 1, wherein the step of preparing CdZnSe QDs comprises:
dissolving cadmium chloride and glutathione in deionized water to obtain a mixed solution A;
dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B;
adjusting the pH value of the mixed solution A and the mixed solution B to 9.0-11.0;
centrifuging selenium powder and sodium borohydride at room temperature until the solution is colorless, so as to obtain a mixed solution C;
mixing the mixed solution C, the mixed solution A and the mixed solution B, and then heating in a water bath to react to obtain a CdZnSe QDs solution;
and separating and purifying the CdZnSe QDs solution to obtain the CdZnSe QDs.
3. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein in the step of dissolving cadmium chloride and glutathione in deionized water to obtain a mixed solution A, the molar ratio of cadmium chloride to glutathione is 1:2.
4. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein in the step of dissolving zinc chloride and glutathione in deionized water to obtain a mixed solution B, the molar ratio of the zinc chloride to the glutathione is 1 (0.5-2).
5. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein the molar ratio of the mixed solution A to the mixed solution B is 1 (0.1-0.2).
6. The method for synthesizing CdZnSe/Mn/ZnS QDs according to claim 2, wherein in the step of adjusting the pH of the mixed solution a and the mixed solution B to 9.0 to 11.0, specifically: and regulating the pH value of the mixed solution A and the mixed solution B to 9.0-11.0 by using sodium hydroxide solution, wherein the concentration of the sodium hydroxide solution is 0.5-5 mol/L.
7. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein in the step of centrifugally reacting selenium powder with sodium borohydride at room temperature until the solution is colorless to obtain a mixed solution C, the molar ratio of the selenium powder to the sodium borohydride is 1:1-4.
8. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein in the step of mixing the mixed solution C, the mixed solution A and the mixed solution B and then heating in a water bath to react, the CdZnSe QDs solution is obtained specifically: and (3) uniformly mixing and stirring the mixed solution C, the mixed solution A and the mixed solution B, transferring the mixture into a water bath kettle, and reacting at 95 ℃ for 30-60 min to obtain the CdZnSe QDs.
9. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 8, wherein CdCl is contained in the mixed solution C, the mixed solution A and the mixed solution B 2 The concentration is 0.001-0.0625 mM.
10. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 2, wherein in the step of separating and purifying the CdZnSe QDs solution to obtain CdZnSe QDs, specifically:
and (3) carrying out centrifugal purification on the CdZnSe QDs solution by using an ultrafiltration tube, removing waste liquid, adding ultrapure water, washing and centrifuging, inversely centrifuging the ultrafiltration tube to obtain CdZnSe QDs, and storing at 4 ℃ for standby, wherein the molecular retention of the ultrafiltration tube is 30KD, the washing and centrifuging times are 4 times, the rotating speed during each centrifuging is 10000r/min, and the centrifuging time is 10min.
11. The method for synthesizing CdZnSe/Mn: znS QDs according to claim 1, wherein the steps of mixing the CdZnSe QDs, manganese chloride, mixed solution B and sodium sulfide solution and then heating in water bath for reaction to obtain the CdZnSe/Mn: znS QDs are specifically as follows:
sequentially adding manganese chloride, mixed solution B and sodium sulfide solution into the CdZnSe QDs, uniformly mixing, transferring to a water bath kettle, and reacting at 80 ℃ for 30min to perform water bath heating reaction to obtain the CdZnSe/Mn ZnS QDs.
12. A CdZnSe/Mn: znS QDs synthesized by the synthesis method of CdZnSe/Mn: znS QDs according to any one of claims 1 to 10.
13. Use of CdZnSe/Mn ZnS QDs according to claim 12 for pH imaging in living cells.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101220275A (en) * | 2008-01-24 | 2008-07-16 | 上海交通大学 | Hydrothermal production method for water-soluble ZnCdSe quantum dot |
| CN104449683A (en) * | 2013-09-23 | 2015-03-25 | 深圳先进技术研究院 | Copper-doped lattice strain quantum dots and preparation method thereof |
| CN107011892A (en) * | 2017-03-02 | 2017-08-04 | 西北大学 | A kind of preparation method and application of Cu Mn codopes ZnS quantum dot solution |
| CN107353903A (en) * | 2017-06-28 | 2017-11-17 | 武汉大学 | A kind of method and its application of synthetic dyestuffs modifying DNA functionalization cadmium content point |
| CN107384368A (en) * | 2017-07-18 | 2017-11-24 | 厦门世纳芯科技有限公司 | A kind of high temperature resistant quantum dot fluorescence material and preparation method thereof |
| CN107643271A (en) * | 2017-08-04 | 2018-01-30 | 华南师范大学 | A kind of salicylic acid Mn doping ZnS quantum points composite nanoparticle Ratiometric fluorescent probe and its preparation method and application |
| CN108048093A (en) * | 2017-12-05 | 2018-05-18 | 湖北大学 | A kind of Ag, Mn adulterate the preparation method of ZnSe and ZnS core-shell structured quantum dot |
| CN109385280A (en) * | 2018-09-28 | 2019-02-26 | 天津工业大学 | Mn2+Adulterate the synthesis in water of CdTe quantum |
| CN109935608A (en) * | 2019-03-04 | 2019-06-25 | 东南大学 | A kind of day blind ultraviolet detection structure and preparation method thereof introducing quantum dot |
| CN111040757A (en) * | 2019-11-01 | 2020-04-21 | 浙江工业大学 | Preparation method and application of ratiometric fluorescent probe for detecting copper ions |
-
2021
- 2021-08-27 CN CN202110996152.1A patent/CN113698935B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101220275A (en) * | 2008-01-24 | 2008-07-16 | 上海交通大学 | Hydrothermal production method for water-soluble ZnCdSe quantum dot |
| CN104449683A (en) * | 2013-09-23 | 2015-03-25 | 深圳先进技术研究院 | Copper-doped lattice strain quantum dots and preparation method thereof |
| CN107011892A (en) * | 2017-03-02 | 2017-08-04 | 西北大学 | A kind of preparation method and application of Cu Mn codopes ZnS quantum dot solution |
| CN107353903A (en) * | 2017-06-28 | 2017-11-17 | 武汉大学 | A kind of method and its application of synthetic dyestuffs modifying DNA functionalization cadmium content point |
| CN107384368A (en) * | 2017-07-18 | 2017-11-24 | 厦门世纳芯科技有限公司 | A kind of high temperature resistant quantum dot fluorescence material and preparation method thereof |
| CN107643271A (en) * | 2017-08-04 | 2018-01-30 | 华南师范大学 | A kind of salicylic acid Mn doping ZnS quantum points composite nanoparticle Ratiometric fluorescent probe and its preparation method and application |
| CN108048093A (en) * | 2017-12-05 | 2018-05-18 | 湖北大学 | A kind of Ag, Mn adulterate the preparation method of ZnSe and ZnS core-shell structured quantum dot |
| CN109385280A (en) * | 2018-09-28 | 2019-02-26 | 天津工业大学 | Mn2+Adulterate the synthesis in water of CdTe quantum |
| CN109935608A (en) * | 2019-03-04 | 2019-06-25 | 东南大学 | A kind of day blind ultraviolet detection structure and preparation method thereof introducing quantum dot |
| CN111040757A (en) * | 2019-11-01 | 2020-04-21 | 浙江工业大学 | Preparation method and application of ratiometric fluorescent probe for detecting copper ions |
Non-Patent Citations (1)
| Title |
|---|
| Relaxation Dynamics of Excited States in Mn2+-Doped ZnCdS (Core)/ZnS (Shell) Quantum Dots Ions in Propylene Carbonate;A. N. Kostrov等;《High Energy Chemistry》;20201120;第54卷(第6期);第462-468页 * |
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