CN112358866A - Orthogonal up-conversion luminescence nano probe, preparation method and application thereof - Google Patents
Orthogonal up-conversion luminescence nano probe, preparation method and application thereof Download PDFInfo
- Publication number
- CN112358866A CN112358866A CN202011284573.3A CN202011284573A CN112358866A CN 112358866 A CN112358866 A CN 112358866A CN 202011284573 A CN202011284573 A CN 202011284573A CN 112358866 A CN112358866 A CN 112358866A
- Authority
- CN
- China
- Prior art keywords
- orthogonal
- conversion
- ucnps
- rdms
- caspase
- 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.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/77—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals
- C09K11/7766—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals containing two or more rare earth metals
- C09K11/7772—Halogenides
- C09K11/7773—Halogenides with alkali or alkaline earth metal
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Physics & Mathematics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Composite Materials (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Manufacturing & Machinery (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses an orthogonal up-conversion nano probe, which is an up-conversion nano composite particle taking up-conversion nano particles as cores, absorbing RdMs and simultaneously modifying a dye Cy5.5 on the surface, namely UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles. The nano probe can respectively emit red light and green light under the excitation of near infrared light with the wavelength of 980nm and 808 nm. The invention also discloses a preparation method of the orthogonal up-conversion nano probe, which comprises the following steps: preparation of a composition having three layersOrthogonal up-conversion nanoparticles of core-shell structure; coating a layer of mesoporous silicon on the surface of the orthogonal up-conversion nano particles for modification to prepare UCNPs @ mSiO2‑NH2A nanoparticle; adsorbing RdMs in the mesoporous pore canal of the particles to obtain UCNPs @ mSiO2@ RdMs; cy5.5 is modified on the surface of the UCNPs @ mSiO2@ RdMs composite nanoparticle to prepare the UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticle. The invention also discloses application of the probe, which can simultaneously detect nitric oxide and Caspase-3 protease through green light quenching and red light recovery, and can prove that NO can promote Caspase-3 to be up-regulated in a cell environment.
Description
Technical Field
The invention relates to the field of biological materials, in particular to an orthogonal up-conversion luminescence-based nano probe, a preparation method thereof and detection in solution and cell environments.
Background
In the prior art, Nitric Oxide (NO) is an important signal molecule in cells, and is widely distributed in various tissues in organisms, and plays an important biological role in cardiovascular and cerebrovascular regulation, nerve regulation, immune regulation and the like. NO as a gas radical can diffuse rapidly in cells and tissues and react with biological targets, and it is very difficult to detect NO formation and migration in vivo. Caspases, an important class of human proteases, play an important role in the initiation and execution of apoptosis (also known as programmed cell death), and are an active regulatory pathway for cell growth and proliferation. Caspases are a broad range of human diseases caused by inappropriate regulation of activity, including neurodegenerative diseases, cardiovascular diseases, immunodeficiency, autoimmune diseases, cancer, and the like. Caspase-3 is a key performer of apoptosis and is involved in both endogenous and exogenous pathways of apoptosis by specific recognition and cleavage of the N-terminus of the Asp-Glu-ValAsp (DEVD) sequence in the substrate protein. Therefore, the real-time monitoring of the NO and Caspase-3 activities in cells and organisms has important significance for the diagnosis and prognosis of diseases and the development of anti-apoptosis drugs.
The down-conversion organic fluorophore, quantum dot or lanthanide chelate currently used is limited by the environmental sensitivity of the fluorescent carrier, such as pH value and hydrophobicity, when the down-conversion organic fluorophore, quantum dot or lanthanide chelate is detected in living cells and living bodies; in addition, the self-luminescence interference, poor membrane permeability and photobleaching of biological samples can be prevented. Thus, the rare earth ion (Ln) was studied3+) Isodoped up-conversion nanoparticles (UCNPs), how to eliminate spontaneous luminescence interference phenomenon of biological samplesThe signal to noise ratio is improved, the sensitivity and the stability are enhanced, and the method becomes a more urgent problem in the aspects of biological detection and cell imaging.
Disclosure of Invention
In order to overcome the above disadvantages of the prior art, the present invention aims to provide an orthogonal upconversion luminescent nanoprobe, which uses an orthogonal upconversion luminescent nanoparticle as a core, wherein the upconversion nanoparticle can emit red light or green light in any one waveband under two different near infrared light excitations simultaneously, i.e. one near infrared excitation light can excite light in one waveband. The invention also aims to provide a preparation method of the orthogonal up-conversion luminescence nano probe.
The invention also aims to provide the application of the orthogonal up-conversion luminescence nano-probe, which can realize the synchronous detection of NO and Caspase-3 based on the fluorescence quenching and fluorescence recovery of the orthogonal up-conversion luminescence nano-particles, and the detection method has the advantages of simple and convenient operation and higher sensitivity.
In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
an orthogonal up-conversion nano probe is characterized in that the probe is an up-conversion nano composite particle which takes up the up-conversion nano particle as a core, adsorbs RdMs and simultaneously modifies a dye Cy5.5 on the surface, namely UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
The UCNPs @ mSiO2@ RdMs @ Cy5.5-pep up-conversion nanoparticles emit any one of red light or green light wave band under excitation of two different near infrared lights, namely, one near infrared excitation light can correspondingly excite light of one wave band: the fluorescent material emits red light under the excitation of 980nm near infrared light and emits green light under the excitation of 808nm near infrared light.
The preparation method of the orthogonal up-conversion nano probe is characterized by comprising the following steps of:
(1) preparing orthogonal up-conversion nanoparticles with a three-layer core-shell structure by adopting a seed crystal method: the nano-particles are prepared by doping Yb, Er and Tm NaYF4The nano particles are cores, and Yb-doped NaYF is wrapped outside the cores4Forming a first shell layer, and then wrapping Yb and Nd doped NaYF outside the first shell layer4Forming a second shell layer to form a Yb, Tm, Er and Nd co-doped core-shell structure to prepare the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd；
(2) Coating a layer of mesoporous silicon on the surface of the orthogonal conversion nano-particles by using Tetraethoxysilane (TEOS), and then using 3-Aminopropyltriethoxysilane (APTES) to carry out UCNPs @ mSiO2Carrying out amination modification on the nano particles to obtain UCNPs @ mSiO2-NH2A nanoparticle;
(3) in UCNPs @ mSiO2-NH2Adsorbing rhodamine B derivative molecules (RdMs) in the mesoporous pore canal of the nano particles to obtain UCNPs @ mSiO2@RdMs；
(4) In UCNPs @ mSiO2@ RdMs composite nanoparticle surface, modifying dye N-hydroxysuccinimide ester (Cy5.5) to obtain UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
The step (1) specifically comprises the following steps:
a. synthesis of NaYF doped with Yb, Tm and Er4Nanoparticles as inner core
10mL of Oleic Acid (OA), 10mL of Octadecene (ODE), 0.84g of NaF solid, and Yb (OAc) were weighed out3 0.0683g、Tm(OAc)3 0.0017g、Er(OAc)30.2752g, adding into a three-neck flask A, magnetically stirring, heating to 110-120 ℃, keeping for 10min until the solid is completely dissolved, and then vacuumizing for 20min to remove water and oxygen; after the reaction is finished, introducing argon, heating to 300 ℃, and reacting for 1 h;
b. wrapping Yb doped NaYF outside the inner core4Forming a first shell layer
Oleic Acid (OA)4mL, Octadecene (ODE)4mL, NaF solid 0.4788g, Yb (OAc) were weighed out separately30.07g of the mixture is added into a three-neck flask B, the mixture is magnetically stirred and heated to 110 to 120 ℃, the mixture is kept for 10min until the solid is completely dissolved, and then the mixture is vacuumized to remove water and oxygen; introducing argon after the reaction is completely finished, heating to 150 ℃, injecting the mixture into the three-neck flask A by using a needle tube at the speed of 0.13mL/min after the reaction in the step a is finished, heating to 300 ℃, and carrying out reverse reactionThe reaction time is 1 h;
c. wrapping Yb and Nd outside the first shell layer to form a second shell layer
Weighing 4mL Oleic Acid (OA), 4mL Octadecene (ODE), and Yb (OAc)3 0.0525g、Nd(OAc)30.4333g, adding into a three-neck flask C, magnetically stirring, heating to 110-120 ℃, keeping for 10min, and then vacuumizing to remove water and oxygen; removing all and then introducing N2After the reaction in the step b is finished, injecting the mixture into the three-neck flask A by using a needle tube at the speed of 0.13mL/min, and reacting for 100min at 300 ℃; cooling to room temperature after the reaction is finished, putting the reaction liquid in the three-neck flask A into a centrifugal tube, and performing centrifugal separation to obtain the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd。
The step (2) specifically comprises the following steps:
0.1g CTAB (cetyl trimethyl ammonium bromide) and 20ml pure water are weighed and stirred at 70 ℃, and then the upconversion nanoparticles obtained in step (1) are added and stirred overnight at room temperature. Adding ultrapure water 40ml, ethanol 6ml and sodium hydroxide 0.0024g, and refluxing at 70 deg.C for 30 min; adding TEOS, refluxing at 70 deg.C for 1.5h, centrifuging and washing; refluxing with ethanol hydrochloride solution to remove CTAB, and preparing water-soluble orthogonal up-conversion nanoparticles UCNPs @ mSiO2(ii) a Taking 5ml of ethanol hydrochloric acid solution and ultrapure water according to the ratio of 19:1, adding 50ul APTES, stirring at room temperature for 5min, adding UCNPs @ mSiO2The particles are stirred for 1h at room temperature, and centrifugally washed to prepare aminated orthogonal up-conversion nanoparticles UCNPs @ mSiO2-NH2。
The step (3) specifically includes the following steps:
the UCNPs @ mSiO prepared in the step (2)2-NH2Dissolving in 5ml ethanol, adding 10mgRdMs, stirring at room temperature for 24h, centrifuging after the reaction is finished, taking precipitate, and alternately washing with ethanol and ultrapure water to prepare UCNPs @ mSiO2@RdMs。
The step (4) specifically includes the following steps:
the UCNPs @ mSiO prepared in the step (3)2@ RdMs are dissolved in PBS, 1mg of cross-linking agent (sulfo-SM CC) is added, shaking is carried out for 1h, centrifugal washing is carried out,dissolving in PBS; adding 1mg of CGDEVDAK (Acp), shaking overnight at room temperature, centrifuging, washing and dissolving in DMF; then adding 1mg of Cy5.5 and 40ul of APTES, stirring for 3h at room temperature, and centrifugally washing to prepare UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
The application of the orthogonal up-conversion luminescence nano probe is characterized in that the application of the orthogonal up-conversion luminescence nano probe in NO detection comprises the following steps:
(1) the reagent DEA/NONONOate, which rapidly releases NO at pH7.4, was selected as the NO donor and DEA/NONONOate was dissolved in NaOH as the NO stock.
(2) Converting the Up-converting nanoparticles UCNPs @ mSiO2The @ RdMs are dissolved in PBS, the solution in the step (1) is added, NO can be continuously released under the buffer action of the PBS, the RdMs and the NO react to be converted into rhodamine B, the absorption of the rhodamine B at 500 nm-600 nm is recovered, the absorption spectrum of the rhodamine B is just coincided with a green light emission region (540nm) of the upconversion nanoparticles under the excitation of 808nm laser, and according to the fluorescence resonance energy transfer, the rhodamine B can quench green light of the upconversion nanoparticles under the excitation of 980nm to finish the detection of NO;
the application of the orthogonal up-conversion luminescence nanoprobe is characterized in that the orthogonal up-conversion luminescence nanoprobe is further applied to detection of Caspase-3, polypeptide with a sequence CGDEVDAK (Acp) can be cut by the Caspase-3, and detection of Caspase-3 protease is carried out through recovery of red light of the nanoprobe under excitation of 980nm, and the method specifically comprises the following steps:
(1) converting the Up-converting nanoparticles UCNPs @ mSiO2The @ RdMs @ Cy5.5-pep is dissolved in PBS, Caspase-3 protease with different amounts is added, and with the increase of the Caspase-3 protease, the recovery effect of the red light peak area of the nano probe under the excitation of 980nm is obviously enhanced, so that the detection of the Caspase-3 protease in the solution of the nano probe under the excitation of 980nm is realized;
the application of the orthogonal up-conversion luminescence nano probe is characterized in that the application of the orthogonal up-conversion luminescence nano probe to synchronous detection of NO and Caspase-3 in a cell system comprises the following steps:
(1) co-incubating the orthogonal up-conversion luminescence nano probe and tumor cells, detecting NO and Caspase-3 in living cells through confocal imaging at 980nm and 808nm and verifying the promotion relation of NO to Caspase-3 up-regulation: specifically, cancer cells, a nano probe and DEA/NONONAte are incubated in a confocal dish, confocal imaging is performed at 808nm and 980nm respectively, NO is detected by quenching green light at 808nm, the cancer cells can be rapidly and effectively killed by the NO, so that the apoptosis enzyme Caspase-3 is up-regulated, and the Caspase-3 is synchronously detected by recovering red light at 980 nm.
The invention has the following beneficial effects:
1. the invention provides an orthogonal up-conversion luminescence nano probe and a preparation method thereof, which adopts a seed crystal method to prepare orthogonal up-conversion nano particles with a three-layer core-shell structure, wherein a kernel is NaYF doped with Yb, Er and Tm4Nanoparticles coated outside the inner core with Yb-doped NaYF4The first shell layer is wrapped with Yb and Nd doped NaYF4A second shell layer which forms a Yb, Tm, Er and Nd co-doped core-shell structure, and finally the orthogonal up-conversion nano-particle NaYF is prepared4:Yb/Tm/Er@NaYF4:Yb@NaYF4Yb/Nd, the up-conversion nano particles emit red light under the excitation of 980nm near infrared light, and emit green light under the excitation of 808nm near infrared light.
The rare earth doped up-conversion luminescent nano material adopted by the invention has the advantages of good photochemical stability and no autofluorescence due to the unique anti-Stokes luminescent property.
The rare earth ion doped up-conversion nanoparticles (UCNPs) provided by the invention can convert Continuous Wave (CW) Near Infrared (NIR) light into visible light with higher energy, and are a novel biocompatible high-efficiency fluorescence donor. The UCNPs have the advantages of larger anti-Stokes displacement, narrow emission bandwidth, low toxicity, less interference by background signals and the like, so that the UCNPs are used as fluorescence donors, the self-luminescence interference phenomenon of a biological sample can be eliminated, the signal-to-noise ratio is improved, the sensitivity and the stability are enhanced, and the advantages in the aspects of biological detection and cell imaging are outstanding.
2. The preparation method of the orthogonal up-conversion nano probe provided by the invention has the advantages of simple process and easy implementation. The prepared nanoprobe has high sensitivity and good selectivity, can realize the detection of nitric oxide and Caspase-3 protease through the fluorescence quenching and fluorescence recovery of the orthogonal up-conversion nanoparticles, is an up-conversion nanoprobe capable of quickly detecting NO and Caspase-3 protease, and mainly realizes the synchronous detection of two detection substances by utilizing the fluorescence resonance energy transfer between two signals of the orthogonal up-conversion nanoparticles and two dyes.
3. The application of the orthogonal transformation nanoprobe provided by the invention can realize the detection of NO and Caspase-3 nanoprobes, UCNPs are used as energy donors, the result can be directly observed under the excitation of near infrared light, and due to the unique optical characteristics of the UCNPs, the background interference is eliminated, the background value is low, the signal is stable, and the sensitivity is greatly improved.
4. The application of the orthogonal up-conversion nanoprobe provided by the invention can directly realize quantitative measurement for the detection of NO and Caspase-3 nanoprobes by virtue of an up-conversion luminescence sensor, and has the advantages of simple and rapid operation, field operation and high result accuracy.
5. By detecting the cell environment of the NO and Caspase-3 nanoprobes, the application of the orthogonal transformation nanoprobe provided by the invention can prove that NO can promote Caspase-3 to be up-regulated, thereby achieving the purpose of synchronously detecting NO and Caspase-3 in living cells. The detection method is based on the fluorescence energy resonance transfer principle and has high sensitivity and high specificity.
Drawings
FIG. 1 is a transmission electron microscope image of orthogonally transformed nanoprobe mesoporous particles of the present invention;
FIG. 2 is a fluorescence spectrum of NO detection with the addition of NOdonor under 808nm excitation by the orthogonal up-conversion nanoprobe of the present invention; curve a is the test result without addition of nodor; curve b is the test result with NOdonor added.
FIG. 3 is a fluorescence spectrum of NO detection with NOdonor added to the orthogonal up-conversion nanoprobe under 980nm excitation, curve a is the test result without NOdonor added; curve b is the test result with NOdonor added.
Detailed Description
Referring to fig. 1 to fig. 3, the technical solution of the present invention is explained in detail by embodiments and drawings.
Example 1:
the orthogonal upconversion nanoprobe provided in this example is an upconversion nanocomposite particle with the upconversion nanoparticle as a core, adsorbing RdMs and simultaneously surface-modifying a dye Cy5.5, i.e., UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles. The UCNPs @ mSiO2@ RdMs @ Cy5.5-pep up-conversion nanoparticles emit any one of red light or green light wave band under excitation of two different near infrared lights, namely, one near infrared excitation light can correspondingly excite light of one wave band: the fluorescent material emits red light under the excitation of 980nm near infrared light and emits green light under the excitation of 808nm near infrared light.
A preparation method of the orthogonal up-conversion nano probe comprises the following steps:
(1) preparing orthogonal up-conversion nanoparticles with a three-layer core-shell structure by adopting a seed crystal method: the nano-particles are prepared by doping Yb, Er and Tm NaYF4The nano particles are cores, and Yb-doped NaYF is wrapped outside the cores4Forming a first shell layer, and then wrapping Yb and Nd doped NaYF outside the first shell layer4Forming a second shell layer to form a Yb, Tm, Er and Nd co-doped core-shell structure to prepare the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd；
(2) Coating a layer of mesoporous silicon on the surface of the orthogonal conversion nano-particles by using Tetraethoxysilane (TEOS), and then using 3-Aminopropyltriethoxysilane (APTES) to carry out UCNPs @ mSiO2Carrying out amination modification on the nano particles to obtain UCNPs @ mSiO2-NH2A nanoparticle;
(3) in UCNPs @ mSiO2-NH2Adsorbing rhodamine B derivative molecules (RdMs) in the mesoporous pore canal of the nano particles to obtain UCNPs @ mSiO2@RdMs；
(4) In UCNPs @ mSiO2@ RdMs composite nanoparticlesSurface modification of dye N-hydroxysuccinimide ester (Cy5.5) to obtain UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
The step (1) specifically comprises the following steps:
a. synthesis of NaYF doped with Yb, Tm and Er4Nanoparticles as inner core
10mL of Oleic Acid (OA), 10mL of Octadecene (ODE), 0.84g of NaF solid, and Yb (OAc) were weighed out3 0.0683g、Tm(OAc)3 0.0017g、Er(OAc)30.2752g, adding into a three-neck flask A, magnetically stirring, heating to 110-120 ℃, keeping for 10min until the solid is completely dissolved, and then vacuumizing for 20min to remove water and oxygen; after the reaction is finished, introducing argon, heating to 300 ℃, and reacting for 1 h;
b. wrapping Yb doped NaYF outside the inner core4Forming a first shell layer
Oleic Acid (OA)4mL, Octadecene (ODE)4mL, NaF solid 0.4788g, Yb (OAc) were weighed out separately30.07g of the mixture is added into a three-neck flask B, the mixture is magnetically stirred and heated to 110 to 120 ℃, the mixture is kept for 10min until the solid is completely dissolved, and then the mixture is vacuumized to remove water and oxygen; after the reaction is finished in the step a, injecting argon into the three-neck flask A at the speed of 0.13mL/min by using a needle tube, heating to 300 ℃, and reacting for 1 h;
c. wrapping Yb and Nd outside the first shell layer to form a second shell layer
Weighing 4mL Oleic Acid (OA), 4mL Octadecene (ODE), and Yb (OAc)3 0.0525g、Nd(OAc)30.4333g, adding into a three-neck flask C, magnetically stirring, heating to 110-120 ℃, keeping for 10min, and then vacuumizing to remove water and oxygen; removing all and then introducing N2After the reaction in the step b is finished, injecting the mixture into the three-neck flask A by using a needle tube at the speed of 0.13mL/min, and reacting for 100min at 300 ℃; cooling to room temperature after the reaction is finished, putting the reaction liquid in the three-neck flask A into a centrifugal tube, and performing centrifugal separation to obtain the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd。
The step (2) specifically comprises the following steps:
0.1g CTAB and 20ml pure water are weighed and stirred at 70 ℃, and then the up-conversion nano particles obtained in the step (1) are added and stirred overnight at room temperature. Adding ultrapure water 40ml, ethanol 6ml and sodium hydroxide 0.0024g, and refluxing at 70 deg.C for 30 min; adding TEOS, refluxing at 70 deg.C for 1.5h, centrifuging and washing; refluxing with ethanol hydrochloride solution to remove CTAB, and preparing water-soluble orthogonal up-conversion nanoparticles UCNPs @ mSiO2(ii) a Taking 5ml according to the proportion of 19:1 of ethanol hydrochloric acid solution and ultrapure water, adding 50ul of APTES, stirring at room temperature for 5min, adding UCNPs @ mSiO2The particles are stirred for 1h at room temperature, and centrifugally washed to prepare aminated orthogonal up-conversion nanoparticles UCNPs @ mSiO2-NH2。
The step (3) specifically includes the following steps:
the UCNPs @ mSiO prepared in the step (2)2-NH2Dissolving in 5ml ethanol, adding 10mgRdMs, stirring at room temperature for 24h, centrifuging after the reaction is finished, taking precipitate, and alternately washing with ethanol and ultrapure water to prepare UCNPs @ mSiO2@RdMs。
The step (4) specifically includes the following steps:
the UCNPs @ mSiO prepared in the step (3)2@ RdMs are dissolved in PBS, added with 1mg of cross-linking agent (sulfo-SM CC) and shaken for 1h, centrifugally washed and dissolved in PBS; adding 1mg of CGDEVDAK (Acp), shaking overnight at room temperature, centrifuging, washing and dissolving in DMF; then adding 1mg of Cy5.5 and 40ul of APTES, stirring for 3h at room temperature, and centrifugally washing to prepare UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
The application of the orthogonal up-conversion luminescence nano probe comprises the following steps when the orthogonal up-conversion luminescence nano probe is applied to detection of NO in a solution:
(1) the reagent DEA/NONONOate, which rapidly releases NO at pH7.4, was selected as the NO donor and DEA/NONONOate was dissolved in NaOH as the NO stock.
(2) Dissolving an orthogonal up-conversion luminescent nano probe into PBS, adding different amounts of the solution obtained in the step (1), releasing NO to convert RdMs into rhodamine B, recovering the absorption of the rhodamine B at 500 nm-600 nm, wherein the absorption spectrum of the rhodamine B is just coincided with a green light emission region (540nm) of the up-conversion nano particle under the excitation of 808nm laser, and according to fluorescence resonance energy transfer, the rhodamine B quenches green light of the up-conversion nano particle under the excitation of 808nm, and the peak of the nano probe at 540nm on the spectrum can be reduced. The more NO donors are added, the quenching effect of the green light peak area under the excitation of 980nm can be obviously enhanced, and the NO detection is completed.
The application of the orthogonal up-conversion luminescence nano probe is further applied to detection of Caspase-3 in a solution. Since Caspase-3 can cleave the polypeptide with the sequence CGDEVDAK (Acp), detection of Caspase-3 protease is carried out by the recovery of red light of the nanoprobe under the excitation of 980nm, which comprises the following steps:
(2) dissolving the orthogonal up-conversion luminescence nanoprobe in PBS, adding different amounts of Caspase-3 protease, and obviously enhancing the recovery effect of the nanoprobe in a red light peak area under the excitation of 980nm along with the increase of the Caspase-3 protease, thereby realizing the detection of the nanoprobe on the Caspase-3 under the excitation of 980 nm;
example 2:
the orthogonal transformation nanoprobe, the preparation method and the application thereof provided by the embodiment are basically the same as those of the embodiment 1, and the difference is that:
the preparation method of the orthogonal up-conversion nano-particles is used for preparing the orthogonal up-conversion nano-particles which excite red light at 980nm and green light at 808nm, and the nano-particles are combined with energy acceptors such as rhodamine (RdB) and Cy5.5 as energy donors to prepare up-conversion nano-probes for detecting NO and Caspase-3 protease in a solution system. The nano probe can synchronously detect NO and Caspase-3 in cells through confocal imaging in a cell system, and further verify the promotion relation of NO to the up-regulation of apoptosis enzyme Caspase-3, and the method comprises the following steps:
1. preparation of UCNPs
(1) The method adopts a seed crystal method to prepare orthogonal up-conversion nano particles with a three-layer core-shell structure, and the kernel of the orthogonal up-conversion nano particles is NaYF doped with Yb, Er and Tm4Nanoparticles coated outside the inner core with Yb-doped NaYF4The first shell layer of (a) is,and Yb and Nd doped NaYF is wrapped outside the first shell layer4A second shell layer which forms a Yb, Tm, Er and Nd co-doped core-shell structure, and finally the orthogonal up-conversion nano-particle NaYF is prepared4:Yb/Tm/Er@NaYF4:Yb@NaYF4Yb/Nd, the up-conversion nano particles emit red light under the excitation of 980nm near infrared light, and emit green light under the excitation of 808nm near infrared light.
The step (1) specifically comprises the following steps:
a. synthesis of NaYF doped with Yb, Tm and Er4Nanoparticles as inner core
10mL of Oleic Acid (OA), 10mL of Octadecene (ODE), 0.84g of NaF solid, and Yb (OAc) were weighed out30.0683g、Tm(OAc)30.0017g、Er(OAc)30.2752g, adding into a three-neck flask A, magnetically stirring, heating to 110-120 ℃, keeping for 10min until the solid is completely dissolved, and then vacuumizing for 20min to remove water and oxygen; after the reaction is finished, introducing argon, heating to 300 ℃, and reacting for 1 h;
b. wrapping Yb doped NaYF outside the inner core4First shell
Oleic Acid (OA)4mL, Octadecene (ODE)4mL, NaF solid 0.4788g, Yb (OAc) were weighed out separately30.07g of the mixture is added into a three-neck flask B, the mixture is magnetically stirred and heated to 110 to 120 ℃, the mixture is kept for 10min until the solid is completely dissolved, and then the mixture is vacuumized to remove water and oxygen; after the reaction is finished in the step a, injecting argon into the three-neck flask A at the speed of 0.13mL/min by using a needle tube, heating to 300 ℃, and reacting for 1 h;
c. a second shell layer of Yb and Nd is wrapped outside the first shell layer
Weighing 4mL Oleic Acid (OA), 4mL Octadecene (ODE), and Yb (OAc)30.0525g、Nd(OAc)30.4333g, adding into a three-neck flask C, magnetically stirring, heating to 110-120 ℃, keeping for 10min, and then vacuumizing to remove water and oxygen; removing all and then introducing N2After the reaction in the step b is finished, injecting the mixture into the three-neck flask A by using a needle tube at the speed of 0.13mL/min, and reacting for 100min at 300 ℃; cooling to room temperature after the reaction is finished, putting the reaction solution in the three-neck flask A into a centrifuge tube,centrifugally separating out the obtained nano particles;
2、UCNPs@mSiO2-NH2the preparation method comprises the following steps:
0.1g CTAB and 20ml pure water are weighed and stirred at 70 ℃, and then the up-conversion nano particles obtained in the step (1) are added and stirred overnight at room temperature. Adding ultrapure water 40ml, ethanol 6ml and sodium hydroxide 0.0024g, and refluxing at 70 deg.C for 30 min; adding TEOS, refluxing at 70 deg.C for 1.5h, centrifuging and washing; refluxing with ethanol hydrochloride solution to remove CTAB, and preparing water-soluble orthogonal up-conversion nanoparticles UCNPs @ mSiO2(ii) a Taking 5ml according to the proportion of 19:1 of ethanol hydrochloric acid solution and ultrapure water, adding 50ul of APTES, stirring at room temperature for 5min, adding UCNPs @ mSiO2The particles are stirred for 1h at room temperature, and centrifugally washed to prepare aminated orthogonal up-conversion nanoparticles UCNPs @ mSiO2-NH2。
3、UCNPs@mSiO2Preparation of @ RdMs comprising the steps of:
taking the UCNPs @ mSiO prepared in the step 22-NH2Dissolving in 5ml ethanol, adding 10mgRdMs, stirring at room temperature for 24h, after the reaction is finished, centrifugally washing to prepare UCNPs @ mSiO2@RdMs。
4、UCNPs@mSiO2Preparation of @ RdMs @ Cy5.5-pep, comprising the steps of:
taking the UCNPs @ mSiO prepared in the step 32@ RdMs are dissolved in PBS, added with 1mg of cross-linking agent (sulfo-SMCC), shaken for 1h, centrifugally washed and dissolved in PBS; adding 1mg of CGDEVDAK (Acp), shaking overnight at room temperature, centrifuging, washing and dissolving in DMF; then adding 1mg of Cy5.5 and 40ul of APTES, stirring for 3h at room temperature, and centrifugally washing to prepare UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
5. The method for detecting NO and Caspase-3 by applying the orthogonal up-conversion luminescence nano probe in a cell system comprises the following steps:
(1) co-incubating the up-conversion nano probe and a tumor cell at 37 ℃, detecting NO and Caspase-3 in a living cell through confocal imaging at 980nm and 808nm, and verifying the promotion relation of NO on Caspase-3 up-regulation. And (3) co-incubating cancer cells, the nanoprobes and DEA/NONONOate with different concentrations in a confocal cuvette, performing confocal imaging at 808nm and 980nm respectively, and detecting NO by quenching green light at 808 nm. The generation of NO can quickly and effectively kill cancer cells, thereby leading the up-regulation of the cell apoptosis enzyme Caspase-3 and the recovery of red light under 980nm to achieve the aim of synchronously detecting Caspase-3,
FIGS. 2 and 3 of the present invention are fluorescence spectra before and after adding NODonor under excitation of 808nm and 980nm, respectively, where curve a is the test result without adding NO DONOR; curve b is the test result with NOdonor added.
The invention takes the detection of NO and Caspase-3 as an example, and realizes the detection of the nitric oxide and Caspase-3 protease through the fluorescence quenching and the fluorescence recovery of the orthogonal up-conversion nanoparticles. As can be seen from the graphs in FIGS. 2 and 3, the nanoprobe can emit red light and green light respectively under the excitation of near infrared light with the wavelength of 980nm and 808nm, simultaneously detect nitric oxide and Caspase-3 protease through green light quenching and red light recovery, and can prove that NO can promote the up-regulation of Caspase-3 in a cell environment. The advantages of the nanoprobe are as follows: the rare earth doped up-conversion luminescent nano material has the advantages of good photochemical stability and no autofluorescence due to the unique anti-Stokes luminescent property; the detection method is based on the fluorescence energy resonance transfer principle, and has the characteristics of high sensitivity and high specificity.
The above description is only exemplary of the present invention and should not be taken as limiting the invention, and any modifications, equivalents, improvements and the like that are made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (7)
1. An orthogonal up-conversion nano probe is characterized in that the probe is an up-conversion nano composite particle which takes up the up-conversion nano particle as a core, adsorbs RdMs and simultaneously modifies a dye Cy5.5 on the surface, namely UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
2. The orthogonal up-conversion nanoprobe of claim 1, wherein the UCNPs @ mSiO2@ RdMs @ cy5.5-pep up-conversion nanoparticies emit either red or green light under two different near infrared light excitations, i.e. one near infrared excitation light can correspondingly excite one wavelength band of light: the fluorescent material emits red light under the excitation of 980nm near infrared light and emits green light under the excitation of 808nm near infrared light.
3. A method for preparing the orthogonal up-conversion nanoprobe according to claim 1 or 2, characterized in that it comprises the following steps:
(1) preparing orthogonal up-conversion nanoparticles with a three-layer core-shell structure by adopting a seed crystal method: the nano-particles are prepared by doping Yb, Er and Tm NaYF4The nano particles are cores, and Yb-doped NaYF is wrapped outside the cores4Forming a first shell layer, and then wrapping Yb and Nd doped NaYF outside the first shell layer4Forming a second shell layer to form a Yb, Tm, Er and Nd co-doped core-shell structure to prepare the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd；
(2) Coating a layer of mesoporous silicon on the surface of the orthogonal conversion nano-particles by using Tetraethoxysilane (TEOS), and then using 3-Aminopropyltriethoxysilane (APTES) to carry out UCNPs @ mSiO2Carrying out amination modification on the nano particles to obtain UCNPs @ mSiO2-NH2A nanoparticle;
(3) in UCNPs @ mSiO2-NH2Adsorbing rhodamine B derivative molecules (RdMs) in the mesoporous pore canal of the nano particles to obtain UCNPs @ mSiO2@RdMs；
(4) In UCNPs @ mSiO2@ RdMs composite nanoparticle surface, modifying dye N-hydroxysuccinimide ester (Cy5.5) to obtain UCNPs @ mSiO2@ RdMs @ Cy5.5-pep nanoparticles.
4. The method for preparing an orthogonal transformation nanoprobe according to claim 3, wherein the step (1) specifically comprises the steps of:
a. synthesis of NaYF doped with Yb, Tm and Er4Nanoparticles as inner core
10mL of Oleic Acid (OA), 10mL of Octadecene (ODE), 0.84g of NaF solid, and Yb (OAc) were weighed out30.0683g、Tm(OAc)30.0017g、Er(OAc)30.2752g, adding into a three-neck flask A, magnetically stirring, heating to 110-120 ℃, keeping for 10min until the solid is completely dissolved, and then vacuumizing for 20min to remove water and oxygen; after the reaction is finished, introducing argon, heating to 300 ℃, and reacting for 1 h;
b. wrapping Yb doped NaYF outside the inner core4Forming a first shell layer
Oleic Acid (OA)4mL, Octadecene (ODE)4mL, NaF solid 0.4788g, Yb (OAc) were weighed out separately30.07g of the mixture is added into a three-neck flask B, the mixture is magnetically stirred and heated to 110 to 120 ℃, the mixture is kept for 10min until the solid is completely dissolved, and then the mixture is vacuumized to remove water and oxygen; after the reaction is finished in the step a, injecting argon into the three-neck flask A at the speed of 0.13mL/min by using a needle tube, heating to 300 ℃, and reacting for 1 h;
c. wrapping Yb and Nd outside the first shell layer to form a second shell layer
Weighing 4mL Oleic Acid (OA), 4mL Octadecene (ODE), and Yb (OAc)30.0525g、Nd(OAc)30.4333g, adding into a three-neck flask C, magnetically stirring, heating to 110-120 ℃, keeping for 10min, and then vacuumizing to remove water and oxygen; removing all and then introducing N2After the reaction in the step b is finished, injecting the mixture into the three-neck flask A by using a needle tube at the speed of 0.13mL/min, and reacting for 100min at 300 ℃; cooling to room temperature after the reaction is finished, putting the reaction liquid in the three-neck flask A into a centrifugal tube, and performing centrifugal separation to obtain the orthogonal up-conversion nano-particle NaYF4:Yb/Tm/Er@NaYF4:Yb@NaYF4:Yb/Nd。
5. Use of the orthogonal up-conversion luminescence nanoprobe according to claim 1 or 2, for performing the detection of NO, comprising the steps of:
a reagent DEA/NONONOate is selected as a donor of NO, the DEA/NONOate is dissolved in an aqueous solution of NaOH to be used as a stock solution of NO, the reagent rapidly releases NO under the condition of pH7.4, RdMs and the NO react to be converted into rhodamine B, the absorption of the rhodamine B at 500 nm-600 nm is recovered, the absorption spectrum of the rhodamine B is just coincided with a green light emission region (540nm) of the upconversion nanoparticles under the excitation of 808nm laser, and the rhodamine B can quench green light of the upconversion nanoparticles under the excitation of 808nm according to fluorescence resonance energy transfer to finish the detection of the NO.
6. The use of the orthogonal up-conversion luminescence nanoprobe according to claim 5, wherein the orthogonal up-conversion luminescence nanoprobe material is further used for detecting Caspase-3, the Caspase-3 cleavage sequence is CGDEVDAK (Acp) polypeptide, and the detection of Caspase-3 is performed by the recovery of red light of the nanoprobe under excitation of 980nm, which comprises the following steps:
the orthogonal up-conversion luminescence nanoprobe is dissolved in PBS, different amounts of Caspase-3 protease are added, and with the increase of Caspase-3 protease, the recovery effect of the nanoprobe in a red light peak area under the excitation of 980nm can be obviously enhanced, so that the detection of Caspase-3 protease by the nanoprobe under the excitation of 980nm is realized.
7. Use of the orthogonal up-conversion luminescence nanoprobe according to claim 6, characterized in that it further comprises the following steps:
co-incubating the orthogonal up-conversion luminescence nano probe and tumor cells, detecting NO and Caspase-3 in living cells through confocal imaging at 980nm and 808nm and verifying the promotion relation of NO to Caspase-3 up-regulation: specifically, cancer cells, a nano probe and DEA/NONONAte are incubated in a confocal dish, confocal imaging is performed at 808nm and 980nm respectively, NO is detected by quenching green light at 808nm, the cancer cells can be rapidly and effectively killed by the NO, so that the apoptosis enzyme Caspase-3 is up-regulated, and the Caspase-3 is synchronously detected by recovering red light at 980 nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011284573.3A CN112358866A (en) | 2020-11-17 | 2020-11-17 | Orthogonal up-conversion luminescence nano probe, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011284573.3A CN112358866A (en) | 2020-11-17 | 2020-11-17 | Orthogonal up-conversion luminescence nano probe, preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112358866A true CN112358866A (en) | 2021-02-12 |
Family
ID=74515050
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011284573.3A Pending CN112358866A (en) | 2020-11-17 | 2020-11-17 | Orthogonal up-conversion luminescence nano probe, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112358866A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115216288A (en) * | 2022-05-26 | 2022-10-21 | 潍坊科技学院 | High-sensitivity double-flux detection immunochromatographic test strip based on double-excitation orthogonal emission up-conversion luminescent nanoparticles |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104592996A (en) * | 2015-01-09 | 2015-05-06 | 上海大学 | Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe |
| CN108152253A (en) * | 2017-09-08 | 2018-06-12 | 东华理工大学 | UCNPs@mSiO2- RBH novel nanos probe synthesis and Pb2+The detection method of ion |
-
2020
- 2020-11-17 CN CN202011284573.3A patent/CN112358866A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104592996A (en) * | 2015-01-09 | 2015-05-06 | 上海大学 | Nano probe for copper ion fluorescence imaging in cells and preparation method for nano probe |
| CN108152253A (en) * | 2017-09-08 | 2018-06-12 | 东华理工大学 | UCNPs@mSiO2- RBH novel nanos probe synthesis and Pb2+The detection method of ion |
Non-Patent Citations (3)
| Title |
|---|
| JING WANG ET AL.,: ""Spectral engineering of lanthanide-doped upconversion nanoparticles and their biosensing applications"", 《MATERIALS CHEMISTRY FRONTIERS》 * |
| XIAOQIAN GE, ET AL.,: ""Controlled optical characteristics of lanthanide doped upconversion nanoparticles for emerging applications"", 《DALTON TRANSACTIONS》 * |
| 孙丽宁等: ""具有红光发射的稀土上转换发光纳米晶及其介孔直接功能化"", 《中国稀土学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115216288A (en) * | 2022-05-26 | 2022-10-21 | 潍坊科技学院 | High-sensitivity double-flux detection immunochromatographic test strip based on double-excitation orthogonal emission up-conversion luminescent nanoparticles |
| CN115216288B (en) * | 2022-05-26 | 2024-04-26 | 潍坊科技学院 | High-sensitivity double-flux detection immunochromatography test strip based on double-excitation orthogonal emission up-conversion luminescence nano-particles |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yao et al. | Upconversion luminescence nanomaterials: A versatile platform for imaging, sensing, and therapy | |
| Feng et al. | Upconversion‐nanophosphor‐based functional nanocomposites | |
| EP1877772B1 (en) | Photoluminescent silica-based sensors and methods of use | |
| Arai et al. | Exploring the use of upconversion nanoparticles in chemical and biological sensors: from surface modifications to point-of-care devices | |
| Hada et al. | Folic acid functionalized gold nanoclusters for enabling targeted fluorescence imaging of human ovarian cancer cells | |
| CN102574677B (en) | Silica nanoparticles incorporating chemiluminescent and absorbing active molecules | |
| Liao et al. | Photoinduced electron transfer of poly (o-phenylenediamine)–Rhodamine B copolymer dots: application in ultrasensitive detection of nitrite in vivo | |
| Li et al. | Strategies for constructing upconversion luminescence nanoprobes to improve signal contrast | |
| CN105482809B (en) | A kind of sulphur hydrogen radical ion nano-probe material and its preparation method and application | |
| Chen et al. | Architecting ultra-bright silanized carbon dots by alleviating the spin-orbit coupling effect: a specific fluorescent nanoprobe to label dead cells | |
| CN111175266A (en) | Construction method and detection method of near-infrared fluorescence biosensor | |
| WO2022116672A1 (en) | Tumor microenvironment response-type off-on upconversion fluorescent probe, preparation method therefor, and application thereof | |
| Ke et al. | Multiplexed intracellular detection based on dual-excitation/dual-emission upconversion nanoprobes | |
| Han et al. | A ratiometric nanoprobe consisting of up-conversion nanoparticles functionalized with cobalt oxyhydroxide for detecting and imaging ascorbic acid | |
| WO2014200164A1 (en) | Composition having ultra-high sensitivity and high selectivity for detecting copper ion and fluorescent chemical sensor | |
| JP6141186B2 (en) | Multiple dye-doped silica nanoparticles featuring highly efficient energy transfer and tunable stalk shift | |
| CN112375561B (en) | Up-conversion fluorescent nanoprobe and application thereof | |
| Wang et al. | Selectively light-up hydrogen peroxide in hypoxic cancer cells with a novel fluorescent probe | |
| CN106947467A (en) | It is a kind of to detect arginic up-conversion luminescence nanosensor material and the application in arginine detection | |
| CN114675026A (en) | Dissolution-enhanced long afterglow luminescence detection method | |
| CN112126428A (en) | Orange fluorescent carbon dot and preparation method and application thereof | |
| Seo et al. | Single-photon-driven up-/down-conversion nanohybrids for in vivo mercury detection and real-time tracking | |
| CN112358866A (en) | Orthogonal up-conversion luminescence nano probe, preparation method and application thereof | |
| CN113025323B (en) | Excitation adjustable double-emission N-doped carbon dot and preparation method and application thereof | |
| Zhu et al. | Lanthanide coordination polymers used for fluorescent ratiometric sensing of H2O2 and glucose |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210212 |