CN108204951A - A kind of photoacoustic imaging probe and its preparation method and application - Google Patents
A kind of photoacoustic imaging probe and its preparation method and application Download PDFInfo
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- CN108204951A CN108204951A CN201611175325.9A CN201611175325A CN108204951A CN 108204951 A CN108204951 A CN 108204951A CN 201611175325 A CN201611175325 A CN 201611175325A CN 108204951 A CN108204951 A CN 108204951A
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- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/1702—Systems in which incident light is modified in accordance with the properties of the material investigated with opto-acoustic detection, e.g. for gases or analysing solids
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Abstract
The present invention proposes a kind of photoacoustic imaging probe and its preparation method and application, by liposome and cyanine dye by being self-assembly of water-soluble nano probe, wherein, cyanine dye has specificly-response to methyl mercury, when there are MeHg in solution+When, nano-probe will be promoted cyclization to form LP hCy7 ' by mercury and the absorption peak strength at 690nm be caused to reduce and the absorption peak strength increase at 860nm, and the signal that accordingly resulting in the photoacoustic signal at 690nm reduces and be located at 860nm significantly increases.MeHg in biological sample is can detect by organism photoacoustic imaging and using PA signal ratio of the LP hCy7 nano-probes at 860nm and 690nm+Content.The photoacoustic imaging probe, simple in structure while preparation process are simple and convenient to operate, the equipment for not needing to complex and expensive, it is easy to accomplish industrialized production.
Description
Technical field
The present invention relates to field of biological detection, particularly relate to a kind of photoacoustic imaging probe and its preparation method and application.
Background technology
Mercury (mercury) is a kind of common heavy metal contaminants, can lead to serious water pollution and soil pollution.Methyl mercury
(MeHg+) be mercury a kind of organic form, and than mercury other inorganic salt forms have stronger toxicity.Methyl mercury is a kind of fat
Soluble substance, therefore it to pass through biomembrane and can cause the damage of organism brain and nervous system, especially to pregnant woman and
Baby is very harmful.Methyl mercury is easy to accumulate among human body by food such as fish.At present, for the detection of methyl mercury
Mainly include gas-chromatography (GC), high performance liquid chromatography (HPLC), Capillary Electrophoresis (CE) coupling specificity detection as atom is glimmering
Light spectrum (AFS), mass spectrum (MS) or inductivity coupled plasma mass spectrometry (ICPMS) etc..However, preceding place of these methods to sample
Manage sufficiently complex, therefore it is a kind of simple, in real time and visualization method is used for the detection of methyl mercury to be badly in need of development, especially raw
Detection in object biopsy sample.
Photoacoustic imaging (PAI) is a kind of a kind of imaging pattern that ultrasonic wave is generated based on light excitation chromophore.With optoacoustic
The development of contrast agent such as organic dyestuff, carbon nanotube, metal-based nano particle etc., photoacoustic imaging technology are widely used in tumour
Imaging, the monitoring of disease therapeuticing effect and reactive oxygen species, PH variations, the detection of enzymatic activity and metal ion etc. field.
Photoacoustic imaging has the advantages that the living imagings such as simple and quick, high sensitivity, not damaged, non-intruding, therefore, using photoacoustic imaging
Methyl mercury in technology detection organism is a kind of ideal detection method.However, detect organism using photoacoustic imaging technology
During interior concrete analysis object, need to develop the spy that there is specificly-response to analyte and photoacoustce signal intensity can be significantly changed
Different photoacoustic contrast agent.At present, the photoacoustic contrast agent that methyl mercury detects in organism is directed to be not yet reported that.
Invention content
The invention discloses a kind of photoacoustic imaging probes and its preparation method and application, are by liposome (LP) and Hua Jingran
(hCy7) is expected by being self-assembly of water-soluble nano probe (LP-hCy7), wherein, hCy7 has spy to methyl mercury (MeHg+)
Opposite sex response, when, there are during MeHg+, LP-hCy7 will be promoted cyclization to form LP-hCy7 ' by mercury and lead to position in solution
Absorption peak strength at 690nm reduces and the absorption peak strength at 860nm increases, and accordingly results at 690nm
Optoacoustic (PA) signal reduce and the PA signals at the 860nm significantly increase.Therefore, it by organism photoacoustic imaging and utilizes
PA signal ratio (PA860/PA690) of the LP-hCy7 nano-probes at 860nm and 690nm can detect MeHg in biological sample+'s
Content.
The technical proposal of the invention is realized in this way：A kind of photoacoustic imaging probe is by liposome (LP) and Hua Jingran
Material passes through the water-soluble nano probe (LP-hCy7) being self-assembly of.
As a preferred embodiment, the cyanine dye is hCy7.
As a preferred embodiment, the diameter of the nano-probe is in 50~100nm.
The photoacoustic imaging probe utilizes the detectable biology of PA signals ratio (PA860/PA690) at 860nm and 690nm
MeHg in sample+Content.
As another object of the present invention, the invention also discloses the preparation method of the photoacoustic imaging probe, including
Self assembly after synthesizing cyanine dye (hCy7) and mixing the cyanine dye (hCy7) of synthesis with liposome (LP).
As a preferred embodiment, include the following steps：
A) synthesis of cyanine dye hCy7：By the compound 1 with below formula
It is dissolved in anhydrous dimethyl formamide (DMF) and is configured to solution, add 1- (2- amino-ethyls) 3- phenylthioureas
And stir under an argon atmosphere, then, DMF solvent is removed, and pass through the sample prepared by silica gel column purification by reducing pressure,
It is spare；
B) preparation of nano-probe (LP-hCy7)：By the hCy7 of preparation and liposome (PEG5000- DSPE) it is dissolved in dichloro
In methane, the solution is then added in into ultrasonic disperse in pure water, organic solvent is removed finally by rotary evaporation and obtains LP-
HCy7 suspension.
As another object of the present invention, inventor is surprisingly found that, the nano-probe has methyl mercury special
Property response, when in tested sample there are the photoacoustic imaging signal that the nano-probe can be significantly changed during methyl mercury, so as to
With by the photoacoustic imaging probe application in detection methyl mercury.
As a preferred embodiment, cyclization is promoted to form LP- by mercury under the conditions of methyl mercury using nano-probe
HCy7 ' is detected.
As a preferred embodiment, the ultraviolet-ray visible absorbing peak intensity using the nano-probe at 550~750nm
It is reduced with the increase of methyl mercury content in solution, and the ultraviolet-ray visible absorbing peak intensity at 800~950nm is with molten
The increase of methyl mercury content in liquid and increase and be detected.
As a preferred embodiment, using photoacoustic imaging signal of the nano-probe at 690nm with solution in first
The increase of base mercury content and slightly reduce, and the photoacoustic imaging signal at 860nm with solution in methyl mercury content increase
And significantly increase and be detected.
As a preferred embodiment, using photoacoustic imaging signal ratio of the nano-probe at 860nm and 690nm with it is molten
Methyl mercury in 0~5 μM of concentration range there is good linear relationship to be detected in liquid, the nano-probe (LP-
HCy7) the photoacoustic imaging signal ratio (PA860/PA690) at 860nm and 690nm has specificity to methyl mercury, will not be by it
Its cation such as Ca2+、Mg2+、Mn2+、Fe2+、Co2+、Ni2+、Zn2+、Cu2+Deng interference, detection is accurate.
The photoacoustic imaging probe of the present invention, simple in structure while preparation process are simple and convenient to operate, and do not need to be multiple
The equipment of miscellaneous costliness, it is easy to accomplish industrialized production.And LP-hCy7 nano-probes disclosed by the invention are to MeHg+With single-minded
And the characteristics of high sensitivity, therefore will have a good application prospect in the detection field of biological sample.
Description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention, for those of ordinary skill in the art, without having to pay creative labor, may be used also
To obtain other attached drawings according to these attached drawings.
Fig. 1 is the TEM figures of synthesis LP-hCy7 nano-probes in embodiment 2；
Fig. 2 is LP-hCy7 nano-probes in embodiment 3 in vitro to methyl mercury specificity ratiometer photo-acoustic detection figure；
Fig. 3 is that LP-hCy7 nano-probes examine methyl mercury specificity ratiometer optoacoustic in zebra fish live body in embodiment 4
Mapping.
Specific embodiment
Below in conjunction with the attached drawing in the embodiment of the present invention, the technical solution in the embodiment of the present invention is carried out clear, complete
Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained every other without creative efforts
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1
As shown in reaction equation (1), 111mg is had to the compound 1 of below formula
(i.e. compound 1 in reaction equation 1) is dissolved in 5mL anhydrous dimethyl formamides (compound 2 in DMF, i.e. reaction equation 1)
In be configured to solution, add 39mg 1- (2- amino-ethyls) 3- phenylthioureas (compound 2 i.e. in reaction equation 1) and in argon
The lower 80 DEG C of stirrings 8h of gas atmosphere.Then, DMF solvent is removed by reducing pressure, and passes through silicagel column (DCM/MeOH 99:1~
15:1)) the prepared sample of purifying, i.e., the compound 3 in reaction equation finally obtain dark blue powder sample hCy7, this method
The yield for preparing cyanine dye hCy7 is 50%.
Reaction equation 1：Wherein,
Embodiment 2
By 1mg cyanine dyes hCy7 and 15mg liposomes (PEG5000- DSPE) it is dissolved in 1mL dichloromethane, then will
The solution is added to ultrasound 120 seconds in 3mL pure water, removes organic solvent finally by rotary evaporation and obtains blue suspension
Liquid contains the LP-hCy7 nano-probes that grain size as shown in Figure 1 is 50~100nm in the solution.
Embodiment 3
LP-hCy7 is scattered in deionized water, is made into the solution of 2mg/mL.Respectively plus different ions solution (Ca2+,
Mg2+、Mn2+、Fe2+、Co2+、Ni2+、Zn2+、Cu2+、MeHg+) 30min is incubated, using deionized water as control, solution is tested respectively and is existed
Photoacoustic image and signal value at 690nm and 860nm, more different groups of photoacoustic signal value ratio PA860/PA690.Test result is such as
Shown in Fig. 2, compared with control group and other ions, MeHg is added in+Photoacoustic signal dramatically increases at solution group 860nm, and 690nm
Locating signal value reduces, so as to which ratio PA860/PA690 be made significantly to increase.
Embodiment 4
Zebra fish is divided into three groups, first group is cultivated in pure water, and second group (dense containing LP-hCy7 nano-probes
Spend for 0.1mg/mL) water in cultivate, third group first is trained in the water containing LP-hCy7 nano-probes (a concentration of 0.1mg/mL)
15min is supported, then continues to cultivate 10min in the water containing methyl mercury.Finally the abdomen of three groups of zebra fish is carried out optoacoustic into
As signal detection.As shown in figure 3, (wherein, a is zebra fish, and the abdomen shade area of coverage is detection position, and b1-b3 is first group of inspection
Survey signal, b4-b5 be second group of detection signal, b7-b9 for third group detect signal, overlay therein for PA690 with
PA860 covering signals are shown) first group of zebra fish at 690nm and 860nm without apparent photoacoustic signal (in Fig. 3 in c tables
Background shown in), second group of zebra fish shows strong photoacoustic signal at 690nm and is shown at 860nm weak
Photoacoustic signal (shown in the LP-hCy7 in Fig. 3 in c tables), photoacoustic signal of the third group zebra fish at 690nm weaken and
(the LP-hCy7+MeHg in Fig. 3 in c tables is remarkably reinforced in photoacoustic signal at 860nm+It is shown).D, which then reflects, in Fig. 3 does not sympathize with
(LP-hCy7 and LP-hCy7+MeHg under shape+) PA860/PA690 ratio, in LP-hCy7+MeHg+Under situation, ratio is drastically
Rise, detection identification marking can be used as.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention god.
Claims (10)
1. a kind of photoacoustic imaging probe, it is characterised in that：To pass through the water solubility that is self-assembly of by liposome and cyanine dye
Nano-probe.
2. photoacoustic imaging probe as described in the appended claim 1, it is characterised in that：The cyanine dye is hCy7.
3. photoacoustic imaging probe as described in the appended claim 1, it is characterised in that：The diameter of the nano-probe is in 50~100nm.
4. the preparation method of photoacoustic imaging probe as described in any in claim 1-3, it is characterised in that：Including synthesizing Hua Jingran
Self assembly after expecting and mixing the cyanine dye of synthesis with liposome.
5. the preparation method of photoacoustic imaging probe as claimed in claim 4, it is characterised in that：Include the following steps：
A) synthesis of cyanine dye：By the compound 1 with below formula
It is dissolved in anhydrous dimethyl formamide and is configured to solution, add 1- (2- amino-ethyls) 3- phenylthioureas and in argon gas gas
It is stirred under atmosphere, then, anhydrous dimethyl formamide solvent is removed, and pass through the sample prepared by silica gel column purification by reducing pressure
Product, it is spare；
B) preparation of nano-probe：By the hCy7 of preparation and liposome dissolving in methylene chloride, then the solution is added in pure
Ultrasonic disperse in water removes organic solvent finally by rotary evaporation and obtains the nano-probe suspension.
6. application of the photoacoustic imaging probe in methyl mercury is detected as described in any in claim 1-3.
7. application of the photoacoustic imaging probe in methyl mercury is detected as recited in claim 6, it is characterised in that：It is visited using nanometer
Needle promotes cyclization formation LP-hCy7 ' to be detected under the conditions of methyl mercury by mercury.
8. application of the photoacoustic imaging probe in methyl mercury is detected as recited in claim 6, it is characterised in that：It is received using described
Ultraviolet-ray visible absorbing peak intensity of the rice probe at 550~750nm with solution in methyl mercury content increase and reduce, and
Ultraviolet-ray visible absorbing peak intensity at 800~950nm with solution in methyl mercury content increase and increase and be detected.
9. application of the photoacoustic imaging probe in methyl mercury is detected as claimed in claim 8, it is characterised in that：It is received using described
Photoacoustic imaging signal of the rice probe at 690nm with solution in methyl mercury content increase and reduce, and the light at 860nm
Acoustic imaging signal increases and is detected with the increase of methyl mercury content in solution.
10. application of the photoacoustic imaging probe in methyl mercury is detected as claimed in claim 9, it is characterised in that：Using described
Photoacoustic imaging signal ratio of the nano-probe at 860nm and 690nm has with methyl mercury in solution in 0~5 μm of concentration range
Some linear relationships are detected.
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| CN112881295A (en) * | 2021-01-15 | 2021-06-01 | 湖南大学 | Ratio photoacoustic nanoprobe based on superoxide anion response and preparation method and application thereof |
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