CN113930235A

CN113930235A - Multi-response nano temperature detection probe, preparation method and application

Info

Publication number: CN113930235A
Application number: CN202111330910.2A
Authority: CN
Inventors: 朱幸俊; 胡倩
Original assignee: ShanghaiTech University
Current assignee: ShanghaiTech University
Priority date: 2021-11-11
Filing date: 2021-11-11
Publication date: 2022-01-14
Anticipated expiration: 2041-11-11
Also published as: CN113930235B

Abstract

The invention discloses a multi-response nano temperature detection probe, a preparation method and application in real-time temperature monitoring. The structural general formula of the temperature detection probe is as follows: NaL_1‑A‑BYb_ATm_BF₄@NaM_1‑CYb_CF₄@NaL_1‑D‑EYb_DNd_EF₄@NaLF₄. The preparation method comprises the following steps: weighing corresponding rare earth and alkali metal chloride according to each layer in sequence, adding a solvent, heating, stirring, dissolving, heating the solution under the protection of nitrogen for reaction, cooling to room temperature, obtaining a solid through centrifugal separation, and washing; finally, ultrasonically dispersing the solid in cyclohexane, adding a methylene chloride solution of distearoyl phosphatidyl ethanolamine-polyethylene glycol, centrifugally separating, and ultrasonically dispersing the solid in water. The invention realizes ratio type luminescence temperature monitoring in the near-infrared second optical window, and has the advantages of no flicker, good stability, low biological tissue autofluorescence, high signal-to-noise ratio, excellent penetration depth and the like.

Description

Multi-response nano temperature detection probe, preparation method and application

Technical Field

The invention relates to a preparation method of a multi-response nano temperature detection probe emitted by a near-infrared second optical window and application of the multi-response nano temperature detection probe in temperature detection in a living body, belonging to the technical field of optical nano probes.

Background

Temperature sensing has profound significance in biomedicine. It helps to reveal the mechanisms of heat generation and conduction, can modulate thermal effects at the cellular and sub-cellular level in a more precise manner, opening up new avenues for explaining disease progression and physiological processes in vivo. However, for the temperature detection in the microscopic level, the conventional measurement method based on the thermocouple, the optical fiber or the thermal imager and other macro devices has the defects that the space precision is not high enough, and only the average temperature value of a certain area can be given, so that the method is not suitable for use. Meanwhile, the method using the thermocouple and the optical fiber also has an invasive problem, and thus, it is necessary to develop a nano/micro material-based thermometer matched to the scale of the observed object. In recent years, several nano thermometers utilizing optical, acoustic and magnetic effects have been reported. The optical nano thermometer has the advantages of high sensitivity, high acquisition speed, real-time continuous measurement, non-invasive and visual detection results, wide range and the like, is more and more concerned by the field, and has very high practical application prospect.

Luminescent materials such as quantum dots, organic fluorophores, and rare earth doped nanoparticles have been used to build optical nanothermometers, but currently some challenges remain in material design. Quantum dots typically contain heavy elements including Pb, Hg, and As, among others, which can present toxicity issues. On the other hand, the organic fluorescent dye has poor light stability, and the emission wavelength thereof is still difficult to extend to the Near Infrared (NIR) region, especially at the near infrared second optical window (NIR-II), i.e. 1000-1700 nm. The rare earth doped luminescent nano particle has the unique advantages of no flicker, excellent light stability, wide emission coverage range, adjustable wavelength and the like, and is an ideal material for an optical nano thermometer. However, the emission wavelength of the current rare earth doped luminescent nano-materials for sensing temperature is usually lower than 1200 nm. The longer wavelengths (>1200nm) in the NIR-II window exhibit better imaging resolution and sensitivity than the shorter wavelengths, and it is therefore of great interest to develop NIR-II nanothermometers with longer operating wavelengths to improve imaging-based thermometry performance.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: a non-invasive optical thermometric material for living body is provided. The material can generate 1-3 near infrared emission bands positioned in 1210-1500nm band under the excitation of 700-1000nm laser, so as to realize the construction of 1-2 optical temperature measurement ratio meters in the same nanometer temperature detection probe, wherein the thermal sensitivity of the temperature measurement ratio meter is 0.6-1.3 percent DEG C^-1. The temperature change in the physiological process of the organism can be monitored in real time by utilizing the obtained nano temperature detection probe in an optical imaging mode. The material has the remarkable characteristics of being capable of generating a plurality of temperature-responsive emission bands positioned in a living body near-infrared second optical window under the excitation of near-infrared light, having good penetration depth and imaging resolution in biological tissues and being used for non-invasive imaging detection of the temperature of a living body.

In order to solve the above technical problems, the present invention provides a multi-response nano temperature detection probe, which is a photoluminescent core-multi shell structure nano material, and the general structural formula of the multi-response nano temperature detection probe is: NaL_1-A-BYb_ATm_BF₄@NaM_1-CYb_CF₄@NaL_1-D- _EYb_DNd_EF₄@NaLF₄Which in turn comprises a first layer NaL from the inside out_1-A-BYb_ATm_BF₄Second layer of NaM_1-CYb_CF₄Third layer NaL_1-D-EYb_DNd_EF₄Fourth layer of NaLF₄The total 4 layers of the structure, wherein M, L is selected from one of Y, Gd elements, A is more than or equal to 0.1 and less than or equal to 0.6, B is more than or equal to 0.005 and less than or equal to 0.03, C is more than or equal to 0.1 and less than or equal to 0.5, D is more than or equal to 0.1 and less than or equal to 0.3, and E is more than or equal to 0.1 and less than or equal to 0.3.

Preferably, the structural formula is NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄Or NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄。

The invention also provides a preparation method of the multi-response nano temperature detection probe, which comprises the following steps:

step 1): according to a first layer NaL_1-A-BYb_ATm_BF₄Weighing corresponding rare earth and alkali metal chloride, adding a solvent, heating and stirring at 120-150 ℃ for 5-30 minutes to dissolve the rare earth and alkali metal chloride to form a uniform solution, and then steaming methanol in an open manner;

step 2): heating the solution obtained in the step 1) to 290-330 ℃ under the protection of nitrogen, reacting for 30-90 minutes, and then cooling to room temperature;

step 3): adding ethanol into the solution obtained in the step 2), obtaining a solid through centrifugal separation, and washing the obtained solid for multiple times by using a mixed solution of ethanol and cyclohexane;

step 4): according to a second layer of NaM_1-CYb_CF₄Weighing corresponding rare earth and alkali metal chloride, adding a solvent, heating and stirring at 120-150 ℃ for 5-30 minutes to dissolve to form a uniform solution, and then steaming methanol in an open manner;

step 5): adding the solid obtained in the step 3) into the solution obtained in the step 4), heating and stirring at 80-100 ℃ for 10-20 minutes, then heating to 290-330 ℃ under the protection of nitrogen, reacting for 30-90 minutes, and then cooling to room temperature;

step 6): adding ethanol into the solution obtained in the step 5), obtaining a solid through centrifugal separation, and washing the obtained solid for multiple times by using a mixed solution of ethanol and cyclohexane;

step 7): according to a third layer NaL_1-D-EYb_DNd_EF₄Weighing corresponding rare earth and alkali metal chloride, adding a solvent, heating and stirring at 120-150 ℃ for 5-30 minutes to dissolve to form a uniform solution, and then steaming methanol in an open manner;

step 8): adding the solid obtained in the step 6) into the solution obtained in the step 7), heating and stirring at 80-100 ℃ for 10-20 minutes, then heating to 290-330 ℃ under the protection of nitrogen, reacting for 30-90 minutes, and then cooling to room temperature;

step 9): adding ethanol into the solution obtained in the step 8), obtaining a solid through centrifugal separation, and washing the obtained solid for multiple times by using a mixed solution of ethanol and cyclohexane;

step 10): according to the fourth layer of NaLF₄Weighing corresponding rare earth and alkali metal chloride, adding a solvent, heating and stirring at 120-150 ℃ for 5-30 minutes to dissolve to form a uniform solution, and then steaming methanol in an open manner;

step 11): adding the solid obtained in the step 9) into the solution obtained in the step 10), heating and stirring at 80-100 ℃ for 10-20 minutes, then heating to 290-330 ℃ under the protection of nitrogen, reacting for 30-90 minutes, and then cooling to room temperature;

step 12): adding ethanol with the same volume into the solution obtained in the step 11), obtaining a solid through centrifugal separation, washing the obtained solid for multiple times by using a mixed solution of ethanol and cyclohexane, and finally ultrasonically dispersing the solid in the cyclohexane;

step 13): adding a dichloromethane solution of distearoyl phosphatidyl ethanolamine-polyethylene glycol with the volume of 1-2 times of that of the cyclohexane solution obtained in the step 12), then carrying out centrifugal separation to obtain a solid, and dispersing the solid in water to obtain the multi-response nano temperature detection probe.

Preferably, in the step 1), the step 4), the step 7) and the step 10), the solvent is at least one of oleic acid and 1-octadecene.

Preferably, in the step 1), the step 4), the step 7) and the step 10), the time for evaporating the methanol in the open air is 30-60 minutes.

Preferably, in the step 3), the step 6), the step 9) and the step 12), the volume ratio of ethanol to cyclohexane in the mixed solution of ethanol and cyclohexane is 1: 1-5: 1, and the obtained solid is washed for 2-3 times.

Preferably, in the step 12), the solid is ultrasonically dispersed in cyclohexane, and the concentration is 10-20 mg/mL.

Preferably, in the step 13), the solid is ultrasonically dispersed in deionized water, and the concentration is 0.1-10 mg/mL.

The invention also provides application of the multi-response nano temperature detection probe, which is used for real-time temperature monitoring and comprises the following steps:

0.1-10mg/mL of aqueous dispersion containing the multi-response nano temperature detection probe, and the using power density of 50-500 mW/cm²Irradiating near-infrared laser with the wavelength of 700-1000nm to realize microscopic temperature rise of the laser in an aqueous solution at 10-90 ℃; under the irradiation of near-infrared laser of 700-1000nm, the integral intensity ratio of different emitted bands shows a linear trend along with the change of temperature, namely I_a/I_bcT + d, wherein I_aFor the integrated fluorescence intensity of one of the emission bands a, I_bThe integral fluorescence intensity of one emission band b, d is a constant, T is temperature, c is the change rate of a ratio meter along with the temperature, c and d are fitted according to a variable temperature curve, and the temperature detection is realized by adopting the mathematical relationship;

injecting the physiological saline dispersion liquid containing the multi-response nano temperature detection probe with the concentration of 0.1-10mg/mL into a focus of a (inflammatory mouse), and after 2-24 hours, using 100-1000 mW/cm according to the guide result of microcosmic temperature monitoring²The 700-1000nm laser irradiates an inflammation area of a mouse, so that the living body real-time and non-invasive temperature change monitoring is carried out on the biological tissue marked with the nano particles.

The invention utilizes rare earth element thulium (Tm) by regulating and controlling the nano material matrix³⁺) Neodymium (Nd)³⁺) The near infrared fine energy level of the probe is used as a monitoring waveband of the optical temperature probe, so that the luminous wavelength of temperature detection is expanded to more than 1200nm, and the tissue penetration depth can be further improved. And the multi-band emission of the fine energy levels has a temperature response function, so that the ratio type luminous temperature detection of multi-channel response can be realized, and the error of single band detection is avoided. By thulium (Tm)³⁺) Neodymium (Nd)³⁺) And gadolinium (Gd)³⁺) The cross relaxation process between can enhance NIR-II emission to optimize the performance of in vivo temperature sensing. Under the excitation of laser with the wavelength of 700-1000nm, the material can generate 1-3 emission bands larger than 1200nm, is used for establishing 1-2 independent ratio type temperature sensing systems, and realizes the independent work of 1-2 temperature detection units in the same system. The preparation of the temperature probe for detecting the temperature of the living body can be realized through simple raw material mixing and heating. The invention is a powerful tool for researching nano-scale heat conduction process and monitoring living body temperature, and makes great contribution to the application of temperature probes of rare earth element luminescent materials in biological imaging, biosensing and other biomedicine.

Compared with the prior art, the invention has the following beneficial effects:

1. the invention can realize ratio type luminescence temperature monitoring in the near-infrared second optical window and has the advantages of no flicker, good stability, low biological tissue autofluorescence, high signal-to-noise ratio, excellent penetration depth and the like.

2. The invention can realize the multi-channel response luminescence temperature measurement, and simultaneously obtain 1-2 optical temperature measurement ratiometers in one system. The method can reduce the systematic error of detection and increase the accuracy in cells, tissues and animals.

Drawings

FIG. 1 is a TEM photograph of example 1;

FIG. 2 is an X-ray powder diffraction analysis of example 1;

FIG. 3 is a Fourier transform infrared spectrum of example 2;

FIG. 4 is a dynamic light scattering analysis of example 2;

FIG. 5 is a Near Infrared (NIR) luminescence spectrum of example 3;

FIG. 6 is a transmission electron microscope photograph of example 4;

FIG. 7 is a Near Infrared (NIR) luminescence spectrum of example 6 in an aqueous solution;

FIG. 8 is a temperature-varying emission spectrum of example 7;

FIG. 9 is a temperature-luminescence calibration curve for example 7, ratioed at 1470nm and 1215nm emissions;

FIG. 10 is a temperature-luminescence correction curve for example 7, ratioed at 1330nm and 1215nm emissions;

FIG. 11 is a temperature-varying emission spectrum of example 8;

FIG. 12 is a temperature-luminescence calibration curve for example 8, ratioed at 1470nm and 1215nm emissions;

FIG. 13 is a temperature-luminescence correction curve for example 8, ratioed at 1330nm and 1215nm emissions;

FIG. 14 shows the temperature imaging test of example 9.

Detailed Description

In order to make the invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings.

Example 1

NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄The synthesis of (2):

1mmol of rare earth element chloride (0.5mmol of GdCl)₃、0.49mmol YbCl₃And 0.01mmol of TmCl₃) Mixed with 10mL of Oleic Acid (OA) and 10mL of 1-octadecene in a 100mL three-necked flask, the solution was heated to 120 ℃ and kept for 30min to obtain a clear solution, and then cooled to 50 ℃. NH dissolved in 11mL of methanol₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, heating the system to 300 ℃, keeping the temperature for 1.5 hours, then cooling to room temperature, adding ethanol with the same volume into the cooled solution, obtaining a solid through centrifugal separation, and adding ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). Weighing 1mmol of rare earth element chloride (0.8mmol YCl)₃，0.2mmol YbCl₃) Adding into a mixed solution of 10mL Oleic Acid (OA) and 10mL Octadecene (ODE), heating to 120 deg.C and maintaining for 30min to obtain a transparent solution, cooling to 50 deg.C, adding the solid obtained by centrifugation, heating and stirring at 80-100 deg.C for 20 min, cooling to room temperature, dissolving in 11mL nailNH in alcohols₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, heating the system to 300 ℃, keeping the temperature for 1.5 hours, then cooling to room temperature, adding ethanol with the same volume into the cooled solution, obtaining a solid through centrifugal separation, and adding ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). Weighing 1mmol of rare earth element chloride (0.65mmol of GdCl)₃，0.1mmol YbCl₃And 0.25mmol of NdCl₃) Adding into a mixed solution of 10mL Oleic Acid (OA) and 10mL Octadecene (ODE), heating to 120 deg.C and maintaining for 30min to obtain a transparent solution, cooling to 50 deg.C, adding the solid obtained by centrifugation, heating and stirring at 80-100 deg.C for 20 min, cooling to room temperature, and dissolving NH in 11mL methanol₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, heating the system to 300 ℃, keeping the temperature for 1.5 hours, then cooling to room temperature, adding ethanol with the same volume into the cooled solution, obtaining a solid through centrifugal separation, and adding ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). After that, except that 1mmol GdCl was added₃In addition, the synthesis process of the core-multishell nanoparticles is almost the same as the previous step. Finally the solid was dispersed ultrasonically in 10mL cyclohexane. The morphology of the finally obtained core-multishell structured nanoparticle is shown in fig. 1, and is a hexagon with an average size of 45.1 nm.

NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄The pure hexagonal phase of the intermediate is shown in figure 2.

Example 2

Distearoyl phosphatidyl ethanolamine-polyethylene glycol modified NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄(NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG) preparation:

to 5mL of CH in which 10mg of distearoylphosphatidylethanolamine-polyethylene glycol was dissolved₂Cl₂10mg cyclohexane dispersed NaGd is added to the solution_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄Stirring for about 30min to uniformly mix the components, removing organic matters in a system by using a rotary evaporation mode, adding 3-5 mL of water for ultrasonic dispersion, obtaining a solid through centrifugal separation, washing the obtained solid with deionized water for three times, and finally ultrasonically dispersing the solid in 1mL of water. NaGd after modification of distearoyl phosphatidyl ethanolamine-polyethylene glycol_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄Transformation of the surface ligand is shown in FIG. 3, NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄The particle size distribution of @ PEG is shown in FIG. 4, and the average particle size is about 72.3nm, and the dispersion in the aqueous phase is good.

Example 3

Determination of NaGd Using fluorescence Spectroscopy_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG near Infrared two-region emission intensity:

the core-multi-shell structure nano material is dispersed in water to prepare 1mL of 10mg/mL solution, the solution is placed in a cuvette, and a 808nm semiconductor laser is used for exciting down-conversion fluorescence, as shown in figure 5.

Example 4

NaYF₄，49％Yb，1％Tm@NaYF₄，20％Yb@NaYF₄，10％Yb，25％Nd@NaYF₄The synthesis of (2):

1mmol of rare earth element chloride (0.5mmol of YCl)₃、0.49mmol YbCl₃And 0.01mmol of TmCl₃) Mixed with 10mL of Oleic Acid (OA) and 10mL of 1-octadecene in a 100mL three-necked flask, the solution was heated to 120 ℃ and kept for 30min to obtain a clear solution, and then cooled to 50 ℃. NH dissolved in 11mL of methanol₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, heating the system to 300 ℃, keeping the temperature for 1.5 hours, then cooling to room temperature, adding ethanol with the same volume into the cooled solution, obtaining a solid through centrifugal separation, and adding ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). Weighing 1mmol of rare earth element chloride (0.8mmol YCl)₃，0.2mmol YbCl₃) Adding the mixture into a mixed solution of 10mL of Oleic Acid (OA) and 10mL of Octadecene (ODE), heating to 120 ℃ and keeping for 30min to obtain a transparent solution, then cooling to 50 ℃, adding the solid obtained by centrifugation before the addition into the solution, and heating and stirring at 80-100 ℃ for 20 min. After cooling to room temperature, the NH dissolved in 11mL of methanol₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, heating the system to 300 ℃, keeping the temperature for 1.5 hours, then cooling to room temperature, adding ethanol with the same volume into the cooled solution, obtaining a solid through centrifugal separation, and adding ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). Weighing 1mmol of rare earth element chloride (0.65mmol YCl)₃，0.1mmol YbCl₃And 0.25mmol of NdCl₃) Adding into a mixed solution of 10mL Oleic Acid (OA) and 10mL Octadecene (ODE), heating to 120 deg.C and maintaining for 30min to obtain a transparent solution, cooling to 50 deg.C, adding the solid obtained by centrifugation, heating and stirring at 80-100 deg.C for 20 min, cooling to room temperature, and dissolving NH in 11mL methanol₄F (3.4mmol) and NaOH (2.5mmol) were added to the solution, followed by degassing at 90 ℃ for 30 minutes to remove methanol. Under the protection of nitrogen, the system is heated to 300 ℃ and kept for 1.5 hours, then cooled to room temperature, and cooled to obtain a solutionThe same volume of ethanol was added and the solid was obtained by centrifugation, then ethanol: the resulting solid was washed three times with a solution of cyclohexane (1:1 v/v). After that, except that 1mmol YCl was added₃In addition, the synthesis process of the Y-based core-multishell nanoparticles is almost the same as that of the previous step. Finally the solid was dispersed ultrasonically in 10mL cyclohexane. The morphology of the finally obtained core-multishell structured nanoparticle is shown in fig. 6 as a hexagon with an average size of 48.4 nm.

Example 5

Distearoyl phosphatidyl ethanolamine-polyethylene glycol modified NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄(NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄@ PEG) preparation:

to 5mL of CH in which 10mg of distearoylphosphatidylethanolamine-polyethylene glycol was dissolved₂Cl₂10mg cyclohexane-dispersed NaY was added to the solution_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄Stirring for about 30min to uniformly mix the components, removing organic matters in a system by using a rotary evaporation mode, adding 3-5 mL of water for ultrasonic dispersion, obtaining a solid through centrifugal separation, washing the obtained solid with deionized water for three times, and finally ultrasonically dispersing the solid in 1mL of water.

Example 6

NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG and NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄Near infrared of @ PEGAnd (3) comparing emission spectra of the two regions:

mixing NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG and NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄@ PEG was dispersed in water to prepare a 10mg/mL solution of 1mL in a cuvette, and then excited to down-convert fluorescence by a 808nm semiconductor laser, as shown in FIG. 7, to obtain NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG and NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄@ PEG near infrared two-domain emission of core-multishell nanostructures.

Example 7

Determination of NaGd using variable temperature fluorescence spectroscopy_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG Standard Curve of fluorescence emission as a function of temperature in Water Dispersion:

mixing NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG was dispersed in water to prepare 1mL of a 10mg/mL solution using 230mW/cm²The solution is continuously irradiated by 808nm laser, and an emission spectrum is collected. The spectra were normalized at 1215nm and, as shown in FIG. 8, the emission intensities at 1470nm and 1330nm were found to increase with increasing temperature. The integrated ratios of the emission intensities at 1470nm and 1215nm, and 1330nm and 1215nm are respectively carried out to obtain the 1470nm wavelength emission peak andthe working curve of the ratio of the 1215nm emission peak to the 1330nm wavelength emission peak to the 1215nm emission peak to the temperature change is shown in FIG. 9, and the working curve of the ratio of the 1330nm wavelength emission peak to the 1215nm emission peak to the temperature change is shown in FIG. 10, and the fluorescence intensity of the fluorescent material shows a linear positive correlation trend along with the temperature.

Example 8

Determination of NaGd using variable temperature fluorescence spectroscopy_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG Standard Curve of fluorescence emission as a function of temperature dispersed in the prosthesis:

mixing NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG was dispersed in the prosthesis to prepare 1mL of a 10mg/mL solution using 233mW/cm²The solution is continuously irradiated by 808nm laser, and an emission spectrum is collected. The spectra were normalized at 1215nm and, as shown in FIG. 11, the emission intensities at 1470nm and 1330nm were found to increase with increasing temperature. The integrated ratios of the 1470nm emission intensity and the 1215nm emission intensity are respectively carried out, the obtained working curve of the ratio of the 1470nm wavelength emission peak to the 1215nm emission peak along with the temperature change is shown in FIG. 12, the working curve of the ratio of the 1330nm wavelength emission peak to the 1215nm emission peak along with the temperature change is shown in FIG. 13, and the fluorescence intensity of the fluorescence-sensitive fluorescent material shows a linear positive correlation trend along with the temperature.

Example 9

NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG is used for in vivo bioimaging and temperature monitoring.

Inducing inflammation in the right hind paw of mouse, which is a normal control group, and dispersing NaGd in physiological saline_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG (10mg/mL, 20. mu.L) was injected into both paws of the mice. One hour later, mice were imaged by NIR-II bio-imaging system. In vivo imaging was obtained at 1215nm and 1470nm wavelengths using 1200nm bandpass and 1400nm long-pass filters, respectively, and the results are shown in FIG. 14, with NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄Temperature imaging given by Probe-Tm of @ PEG shows a rise in right paw temperature with an average temperature of 39.6 ℃ and an average temperature of 37.0 ℃ in the left paw, demonstrating NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄@ PEG enables the detection and visualization of temperature in the diagnosis of living diseases and the monitoring of physiological processes.

Claims

1. A multi-response nanometer temperature detection probe is characterized in that the probe is a photoluminescent core-multi-shell structure nanometer material, and the structural general formula of the probe is as follows:

NaL_1-A-BYb_ATm_BF₄@NaM_1-CYb_CF₄@NaL_1-D-EYb_DNd_EF₄@NaLF₄which in turn comprises a first layer NaL from the inside out_1-A-BYb_ATm_BF₄Second layer of NaM_1-CYb_CF₄Third layer NaL_1-D-EYb_DNd_EF₄Fourth layer of NaLF₄The total 4 layers of the structure, wherein M, L is selected from one of Y, Gd elements, A is more than or equal to 0.1 and less than or equal to 0.6, B is more than or equal to 0.005 and less than or equal to 0.03, C is more than or equal to 0.1 and less than or equal to 0.5, D is more than or equal to 0.1 and less than or equal to 0.3, and E is more than or equal to 0.1 and less than or equal to 0.3.

2. The multi-response nano temperature detection probe of claim 1, wherein the structural formula is NaY_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaY_0.65Yb_0.1Nd_0.25F₄@NaYF₄Or NaGd_0.5Yb_0.49Tm_0.01F₄@NaY_0.8Yb_0.2F₄@NaGd_0.65Yb_0.1Nd_0.25F₄@NaGdF₄。

3. The method for preparing a multi-response nano temperature detection probe according to claim 1 or 2, characterized by comprising the following steps:

step 7): according to a third layer NaL_1-D-EYb_DNd_EF₄Weighing corresponding rare earth and alkali metal chloride, adding a solvent, and heating at 120-150 DEG CStirring for 5-30 minutes, dissolving to form a uniform solution, and then steaming methanol in an open mouth;

4. The method for preparing the multi-response nano temperature detection probe according to claim 3, wherein in the steps 1), 4), 7) and 10), the solvent is at least one of oleic acid and 1-octadecene.

5. The method for preparing the multi-response nano temperature detection probe according to claim 3, wherein in the steps 1), 4), 7) and 10), the time for evaporating methanol in an open environment is 30-60 minutes.

6. The method for preparing the multi-response nano temperature detection probe according to claim 3, wherein in the step 3), the step 6), the step 9) and the step 12), the volume ratio of ethanol to cyclohexane in the mixed solution of ethanol and cyclohexane is 1: 1-5: 1, and the obtained solid is washed for 2-3 times.

7. The method for preparing the multi-response nano temperature detection probe according to claim 3, wherein in the step 12), the solid is ultrasonically dispersed in cyclohexane with a concentration of 10-20 mg/mL.

8. The method for preparing the multi-response nano temperature detection probe according to claim 3, wherein in the step 13), the solid is ultrasonically dispersed in deionized water to a concentration of 0.1-10 mg/mL.

9. Use of the multi-response nano temperature detection probe of claim 1 or 2 for real-time temperature monitoring, comprising the steps of:

injecting the physiological saline dispersion liquid containing the multi-response nano temperature detection probe with the concentration of 0.1-10mg/mL into the focus, and after 2-24 hours, injecting the dispersion liquid according to the micro-scaleObserving the guide result of temperature monitoring, using 100-1000 mW/cm²The 700-1000nm laser irradiates an inflammation area of a mouse, so that the living body real-time and non-invasive temperature change monitoring is carried out on the biological tissue marked with the nano particles.