CN104117075A - Gold nanorod multifunctional probe-based nuclide-cerenkov luminescence-CT multi-mode imaging method - Google Patents

Gold nanorod multifunctional probe-based nuclide-cerenkov luminescence-CT multi-mode imaging method Download PDF

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CN104117075A
CN104117075A CN201410370072.5A CN201410370072A CN104117075A CN 104117075 A CN104117075 A CN 104117075A CN 201410370072 A CN201410370072 A CN 201410370072A CN 104117075 A CN104117075 A CN 104117075A
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gold nanorods
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CN104117075B (en
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屈晓超
梁敬宁
王彦然
李蕾
李奕辰
梁继民
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Xidian University
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Abstract

The invention discloses a gold nanorod multifunctional probe-based nuclide-cerenkov luminescence-CT multi-mode imaging method, which applies a <68>Ga-AuNRs-RGD multifunctional molecular probe to the field of multi-mode molecular imaging. The caudal vein of a tumor-bearing mouse is injected with the <68>Ga-AuNRs-RGD multifunctional molecular probe, nuclide-CT imaging is performed by a PET/CT imaging system, cerenkov luminescence-CT imaging is performed by a CLI/CT (cerenkov luminescence imaging/computed tomography) system, and a PET/CT image serving as a source image and a CLI/CT image serving as a target image are fused by means of grey rectification to obtain a nuclide-cerenkov luminescence-CT multi-mode imaging display image based on the <68>Ga-AuNRs-RGD multifunctional molecular probe. According to the method, the <68>Ga-AuNRs-RGD multifunctional molecular probe is applied to multi-mode molecular imaging, has the advantages of increasing optical signal intensity, improving imaging resolution, improving imaging depth and the like, and has the characteristics of excellent dispersion, low toxicity, high living cell membrane permeability and the like.

Description

Nucleic-Cherenkov based on gold nanorods multiprobe is luminous-CT multi-modality imaging method
Technical field
The present invention relates to biomedical imaging field, be specifically related to a kind of nucleic-Cherenkov based on gold nanorods multiprobe luminous-CT multi-modality imaging method.
Background technology
The development of nanotechnology impels a collection of birth with character nano materials such as uniqueness is optical, electrical, magnetic, and they are widely used in the diagnosis and treatment research of tumor.Noble metal nanometer material, especially gold nano grain, its adjustable for height optical characteristics can strengthen the absorption to visible ray and near infrared light greatly, the optical property that it is good and surface chemistry ability can be carried out accurately location and efficient treatment to tumor tissues, are expected to become the new tool of cancer clinical treatment.
At present, mostly be at the gold nano grain of imaging field application the gold nano grain that size homogeneous, colloidal stability are good, be less than 50nm, these nano-particle are spherical in shape, shell dress, bar-shaped, cage shape use aspect the different kinds of molecules imaging techniques such as CT acoustooptics MRI.Gold nano grain can extend the time of staying in blood circulation effectively, has not only extended time of developing, and its extremely low cytotoxicity has also reduced the toxic and side effects to kidney.The gold nano grain easily surface chemistry affinity of control can be coupled with some specific antibody proteins, prepares the nano-probe of functionalization.Have any different in the expressed receptor of crossing of ordinary cells in malignant cell surface, energy specific binding is with the gold nano-material of corresponding protein molecule.Integrin alpha vβ 3high expressed on kinds of tumor cells surface and new vessels endotheliocyte, but in mature blood endothelial cell and most normal organ system α vβ 3do not express or the seldom expression of amount.Rgd peptide is the small peptide that a class contains arginine-glycine-aspartic acid (Arg-Gly-Asp), by α vβ 3as the binding site of tumor, by RGD and α vβ 3specific binding, gold nanorods is combined with tumor cell specific.Gold nanorods (Gold nanorods, AuNRs) is combined with RGD, makes the selectively targeted tumor cell of gold nanorods energy.At tumor locus, the gold nano-material X ray of can decaying well, carries out CT imaging as CT contrast agent, obtains the CT image of tumor locus.
Gold nano optical imagery is very limited in viviperception and clinical conversion, and a lot of research teams attempt by other imaging molecular probes, as nucleic probe 64cu and 111in, optical probe ICG, magnetic resonance probe Fe 3o 4build multifunctional nano probe Deng labelling gold nano-material, carry out living imaging and the treatment research of gold nano target tumor.Radio nuclide imaging is applied to the details that can obtain focus in oncotherapy.But radio nuclide imaging, as highly sensitive in PET, SPECT imaging, not to be subject to investigation depth restriction, but its apparatus expensive, spatial resolution is low.At present, in the diagnosis of gold nano target tumor and treatment research, associating radio nuclide imaging and optical image technology development multifunctional nano probe, be embodied as picture and treat integratedly, in following biomedical research, has great significance.Cerenkov effect is found in 1934 by scientist Cerenkov, is referred to its part energy to be converted into when charged particle moves in medium with superlight speed visible ray and the near infrared light of 400-900nm wave band.Radioisotopic Cherenkov's luminescence imaging (Cerenkov Luminescence Imaging, CLI) is the imaging that utilizes the light of nucleic Cerenkov effect generation to carry out.Cherenkov's luminescence imaging has advantages of many optical imageries, such as highly sensitive, cheap, imaging time is short, be easy to carry out etc.The more important thing is based on Cerenkov effect, can carry out respectively optical imagery and radio nuclide imaging for same isotope labeling molecular probe, obviously be different from the multi-modality imaging that traditional multiple labelling molecular probe carries out, more easily keep the biological activity of molecular probe, and in body based on same molecular probe, bioprocess carries out respectively optics and radio nuclide imaging, is conducive to carry out image co-registration research.The high energy charged particles producing due to multiple radionuclide all meets Cerenkov effect occurrence condition, can produce the optical signalling that can be collected by high-sensitive optical imaging apparatus, therefore can utilize existing isotope labeling molecular probe to carry out clinical optics video picture, can overcome optical dye toxicity and be difficult to use in more greatly clinical deficiency.
Single image mode cannot provide enough comprehensively information for medical diagnosis on disease.In order to realize early diagnosis and the accurate treatment to major disease, obtain sensitiveer, more accurately, more fully the multi-mode molecule imaging of physiological and pathological information becomes focus and the development trend of molecular image development in conjunction with the advantage of different imaging patterns.By the multi-modality imaging of CT, nucleic, Cherenkov's imaging combination, between each mode, can mutually make up the inferior position of imaging separately, and the advantage of utilizing imaging separately obtains the details of tumor locus, for diagnosing and treat the more comprehensive and detailed information that provides.
Summary of the invention
For the deficiencies in the prior art, the object of the invention is, by a kind of special gold nanorods multiprobe, to utilize probe to carry out multi-modality imaging to animal model for tumour.First utilize RGD to tumor vascular targeting, multiprobe specificity is gathered in tumor locus, and gold nanorods carries out CT imaging as CT contrast agent, then passes through radionuclide 68ga carries out radio nuclide imaging and Cherenkov's luminescence imaging.
For above-mentioned purpose, the present invention adopts following technical scheme:
Step 1, the preparation of gold nanorods polyfunctional molecule probe:
Step 1.1, prepares gold nanorods (AuNRs)
Cetyl trimethyl ammonium bromide (CTAB) and gold chloride are mixed, add sodium borohydride, then rapid stirring, obtains crystal seed; Then in the seeded growth liquid that contains silver nitrate, gold chloride, CTAB and ascorbic acid, add gained crystal seed, after leaving standstill, obtained the gold nanorods of specific draw ratio;
Step 1.2, preparation 68ga-AuNRs-RGD polyfunctional molecule probe
Adopt small-molecular peptides and the radiosiotope of arginine-glycine-aspartic acid (RGD) tripeptide sequence 68ga carrys out labelling gold nanorods.
First by difunctional Polyethylene Glycol (OPPS-PEG2K-NHS) bridging agent of excessive different molecular weight respectively with arginine-glycine-aspartic acid (RGD) and 1,4,7,10-tetraazacyclododecanand-1,4,7,10-tetrabasic carboxylic acid (DOTA) mixes, under room temperature, spend the night, form stable bridging agent OPPS-PEG-RGD and OPPS-PEG-DOTA.Then two kinds of bridging agents are mixed with the ratio of 2500:2500:1 with gold nanorods, by the combination of adjacent two thiopyridines bases (OPSS), the stable gold nanorods surface that is connected to of RGD and DOTA.Finally, radiotropism isotope in certain proportion 68in Ga, add the stable gold nanorods coalition being connected of RGD and DOTA, mix, react 20 minutes, make 68ga-AuNRs-RGD polyfunctional molecule probe.
It should be noted that, step 1.2 68the molecular weight difference of difunctional Polyethylene Glycol (OPPS-PEG-NHS) bridging agent in the preparation process of Ga-AuNRs-RGD polyfunctional molecule probe, the OPPS-PEG-NHS molecular weight being connected with RGD is 2000, and the OPPS-PEG-NHS molecular weight being connected with DOTA is 5000.
Step 2, 68the Performance Detection of Ga-AuNRs-RGD polyfunctional molecule probe:
Step 2.1, 68ga-AuNRs-RGD polyfunctional molecule probe stability test
Adopt 10%NaCl solution to detect 68ga-AuNRs-RGD polyfunctional molecule probe stability.By same concentrations 68ga-AuNRs-RGD polyfunctional molecule probe and gold nanorods are resuspended to respectively in 10% NaCl solution of equivalent, then carry out UVs spectral detection, relatively both stability differences.
Step 2.2, 68ga-AuNRs-RGD polyfunctional molecule fluorescence probe quantum efficiency and extracorporeal receptor binding analysis
First, measure 68cherenkov's photo-quantum efficiency of Ga-AuNRs-RGD polyfunctional molecule probe: more different 68same concentration under Ga radioactivity 68cherenkov's flashlight quantum efficiency of Ga-AuNRs-RGD polyfunctional molecule probe, optimization Test parameter, obtains the optimized fluorescence enhancement mode of luminous efficiency 68ga-AuNRs-RGD polyfunctional molecule probe;
Secondly, measure 68the biology performance of Ga-AuNRs-RGD polyfunctional molecule probe: adopt 68the AuNRs-RGD molecular probe of Ga labelling is as radioligand, and unlabelled AuNRs-RGD molecular probe is non-marked part, analyzes α vβ 3the glioma U87 cell of the expression of receptor positive carries out receptors ligand in conjunction with test.Two kinds of parts are combined with suspension cell, detect their radioactive dosage and luminous efficiency.
Step 3, tumor bearing nude mice Animal Model:
Choose glioma cell U87, cultured cell in vitro, in the right shoulder of nude mice place subcutaneous injection of tumor cells several, build Subcutaneous tumor model, after 2-3 week, carry out multi-modality imaging detection;
It should be noted that, after 2-3 week, tumor model diameter carries out multi-modality imaging detection after being greater than 0.5cm again.
Step 4, 68the multi-modality imaging of Ga-AuNRs-RGD polyfunctional molecule probe:
Step 4.1, PET/CT image data acquiring
Adopt PET/CT system to carry out PET/CT data acquisition, mouse tail vein injection to tumor bearing nude mice 68after Ga-AuNRs-RGD polyfunctional molecule probe, carry out PET/CT scanning at different time points, obtain nucleic-CT view data of different time points;
Step 4.2, CLI/CT image data acquiring
Adopt CLI/CT imaging system to carry out CLI/CT data acquisition, mouse tail vein injection to described tumor bearing nude mice 68after Ga-AuNRs-RGD polyfunctional molecule probe, under airtight dark situation, obtain Cherenkov's luminous signal and the CT view data of multiple spectrum, multiple visual angles toy body surface at different time points.
Step 4.3, PET/CT and CLI/CT fusing image data
The CLI/CT that step 4.2 is drawn is as target image, and the PET/CT that step 4.1 draws is as source images, and through pretreatment such as image denoising, enhancings, image is cut apart and carried out feature extraction, and mutual information image registration, based on the image co-registration of image pixel, has just obtained 68ga-AuNRs-RGD polyfunctional molecule probe target to luminous, the CT multi-modality imaging video picture figure of nucleic, the Cherenkov of tumor region.
It should be noted that, in step 4, the α of the RGD target tumor blood vessel on gold nanorods surface vβ 3receptor, carries out specific binding to tumor.
It should be noted that, in step 4, based on the same molecular probe radionuclide in gold nanorods surface 68in the body of Ga, bioprocess carries out respectively optics and radio nuclide imaging, is conducive to carry out image co-registration research.
The present invention adopts 68ga-AuNRs-RGD molecular probe, tumor bearing nude mice is carried out to CT, nucleic, the luminous multi-modality imaging of Cherenkov following significantly beneficial effect:
1. adopt gold nanorods to replace the contrast agent of iodide as CT imaging, can increase significantly imaging time, have no side effect, greatly reduced the dosage of X-ray, and improved spatial resolution;
2. adopt RGD target tumor tissue, by the integrin alpha in RGD and tumor neogenetic blood vessels vβ 3specific binding, make tail vein injection 68Ga-AuNRs-RGD molecular probe can be gathered in preferably tumor locus;
3. adopt a kind of radioelement labelling gold nano molecular probe to complete nucleic and Cherenkov's luminescence imaging;
4. adopt nucleic-Cherenkov luminous-CT multi-modality imaging, nucleic and Cherenkov's luminescence imaging carry out functional imaging to tumor locus, improve imaging depth, CT obtains three distributed intelligences of tumor in Mice Body.
Brief description of the drawings
Fig. 1 is flow chart of the present invention;
Fig. 2 is the transmission electron microscope picture of gold nanorods;
Fig. 3 be gold nanorods and 68the ultraviolet-visible spectrophotometer testing result figure of Ga-AuNRs-RGD polyfunctional molecule probe;
Fig. 4 is 68the Detection of Stability result figure of Ga-AuNRs-RGD polyfunctional molecule probe and gold nanorods.
Detailed description of the invention
Below with reference to accompanying drawing, the invention will be further described, it should be noted that, the present embodiment, taking the technical program as prerequisite, provides detailed embodiment and implementation step, but is not limited to the present embodiment.
As shown in Figure 1, described nucleic-Cherenkov based on gold nanorods multiprobe luminous-CT multi-modality imaging method comprises the steps:
The preparation of gold nanorods (AuNRs):
In the time that temperature is 27-30 DEG C, to 5ml, in the CTAB of 0.2M, add 5ml, the gold chloride of 0.5mM, slowly stirs, and now solution is faint yellow.Then slow, dropwise add 0.6ml, the sodium borohydride of 0.01M freezing point, rapid stirring 2min, now solution has the light yellow light brown that becomes, and finally becomes claret, and crystal seed generates.At 30 DEG C, preserve, in order to follow-up test.
In lucifuge situation, to 5ml, in the CTAB of 0.2M, add 0.25ml, and then the silver nitrate solution of 4mM, then add 5ml, the gold chloride of 1mM, now solution is yellow, add afterwards 70ul, the ascorbic acid of 0.0788M, finally adds the gained crystal seed of 12ul, lucifuge at 30 DEG C, leave standstill 3h, obtain granule gold nanorods comparatively uniformly, the transmission electron microscope picture of gold nanorods as shown in Figure 2.
68the preparation of Ga-AuNRs-RGD polyfunctional molecule probe:
RGD and the DOTA solution of PBS buffer (PH=7.4) preparation 500ul (1mg/ml) then add 8.82 × 10 respectively in RGD and DOTA solution -2the EDC of mmol and NHS, activate the carboxyl of RGD and DOTA by amidation process, after reacting 30min under room temperature, in RGD solution and DOTA solution, add respectively 30mg OPPS-PEG-NHS (MW=2K) and OPPS-PEG-NHS (MW=5K), finally at 4 DEG C, react 6h, the carboxyl after activation is combined with the amino of double-functional group and has been generated two kinds of bridging agent OPPS-PEG-RGD and OPPS-PEG-DOTA.Then two kinds of each 500ul of bridging agent are joined to 2ml, in the gold nanorods solution of 300ug/ml, middling speed stirs, at 25 DEG C, react 24h, by OPPS, RGD and DOTA are stablized and be connected to gold nanorods surface, then centrifugal 10min under 11000r, removes unnecessary unreacted bridging agent, precipitation is resuspended in the deionized water of 1ml, has obtained RGD targeting gold nanorods.
Adopt 68ga radionuclide carries out labelling gold nanorods, to carry out nucleic and Cherenkov's luminescence imaging, first stable to RGD and the DOTA pH value that is connected gold nanorods coalition solution is adjusted to 3.5-4.0, then adds 68ga, both mix with the ratio of 100:1, and 90 DEG C of lucifuges are reacted 20 minutes.The chelating agen DOTA on gold nanorods surface is for label isotope 68ga, the molecular weight of change Polyethylene Glycol (PEG), can control 68ga and gold nanorods surface be coupled distance, obtain 68ga-AuNRs-RGD polyfunctional molecule probe.In preparation process, the UV, visible light spectrophotometer testing result of each step products as shown in Figure 3.
68the stability test of Ga-AuNRs-RGD molecular probe:
Adopt the monodispersity of high level salt solution test gold nanorods material, get concentration and be all 600ug/ml's 68ga-AuNRs-RGD and the each 100ul of AuNRs solution, be resuspended to respectively in 10% the NaCl solution of 1ml, and two kinds of mixed solutions leave standstill 24h at 4 DEG C, then use ultraviolet-visible spectrophotometer to carry out wave spectrum detection, as shown in Figure 4, the wave spectrum obtaining with 68the aqueous solution wave spectrum of Ga-AuNRs-RGD and AuNRs compares, and obtains 68the stability test result of Ga-AuNRs-RGD molecular probe.
68ga-AuNRs-RGD polyfunctional molecule fluorescence quantum efficiency and extracorporeal receptor binding analysis:
First, measure 68cherenkov's photo-quantum efficiency of Ga-AuNRs-RGD polyfunctional molecule probe: adopt PET system acquisition difference 68under Ga radioactivity 68the nucleic signal of Ga-AuNRs-RGD molecular probe is selected area-of-interest on nuclide chart picture, adopts Living Image3.2 software (IVIS Kinetic, Caliper Life Sciences) can obtain the intensity of optical signalling.This patent is found Cherenkov's optics signal and radioactivity linear correlation.Optimization Test parameter, obtains the optimized fluorescence enhancement mode of luminous efficiency 68ga-AuNRs-RGD polyfunctional molecule probe.Secondly, measure 68the biology performance of Ga-AuNRs-RGD polyfunctional molecule probe: adopt α vβ 3the glioma U87 cell of the expression of receptor positive carries out receptors ligand in conjunction with test.Preparation number of cells identical U87 cell suspending liquid is labeled as A, B and organizes each 1ml, then get Au concentration and be 600ug/ml AuNRs and 68it is 0.1nM that Ga-AuNRs-RGD solution (filtration sterilization) is diluted to Au concentration with cell culture medium, respectively gets 100ul, is added to respectively in A, B group cell suspending liquid, cultivates 30 minutes in cell culture incubator.A, B group cell suspending liquid is carried out to nucleic and Cherenkov's luminescence imaging, detect their radioactive dosage and luminous efficiency.
PET/CT image data acquiring:
Adopt nanoScan PET/CT, Mediso, Medical Imaging Systems system is carried out PET/CT image data acquiring to tumor bearing nude mice, this system by a pair of relative to vertical PET detector, X ray emitter, X ray flat panel detector, toy support with rotate translation stage and form, setting image acquisition voltage is 55kv, collection electric current is 146uA, and image acquisition angle is 240.Give tumor-bearing mice tail vein injection 100ul, concentration is 4mg/ml 68after Ga-AuNRs-RGD polyfunctional molecule probe, respectively at 1,3,6,12,24h carries out PET video picture, observes radiopharmaceutical distribution, metabolism and the dynamic change at tumor uptake in vivo.Carry out CT scan, for image registration, fusion provide details simultaneously.
CLI/CT image data acquiring:
Adopt bimodal ZKKS-Direct3D molecular imaging system to carry out CLI/CT image data acquiring to tumor bearing nude mice, this system is made up of Cherenkov's luminescence imaging system and Micro-CT system.Cherenkov's luminescence imaging system is made up of highly sensitive refrigeration ccd detector, scan-type spectrometer device, filter lens group.Micro-CT system is formed and is formed by X ray emitter, X ray flat panel detector, toy support and rotation translation stage.Setting image acquisition voltage is 50kv, and collection electric current is 10mA, 360 ° of image acquisition.Give tumor-bearing mice tail vein injection 100ul, concentration is 4mg/ml 68after Ga-AuNRs-RGD polyfunctional molecule probe, under completely airtight dark situation, before highly sensitive refrigeration CCD, place the band pass filter of one group of different-waveband, Cherenkov's luminous signal of toy body surface is surveyed at multiple visual angles, obtain the White-light image with stent markers dot information, so that influence of noises such as DC-offset corrections simultaneously.Utilize spectrometer device to survey in the enterprising line scanning of certain wave band body surface luminous signal, obtain the spectral distribution of this wave band luminous signal.Carry out CT scan simultaneously, obtain three information of tumor locus, for image registration, fusion provide details.
PET/CT and CLI/CT image co-registration:
The PET/CT obtaining and two kinds of view data of CLI/CT are carried out to denoising, the pretreatment such as enhancing.Unified two kinds of data formats, the image parameters such as image size and resolution.Using mouse tumor region as area-of-interest, select specific segmentation threshold to carry out manual cutting apart to image, draw the geometric properties of image, then optimize the similarity of PET/CT and two kinds of images of CLI/CT using PET/CT as source images, CLI/CT is as target image, use gray scale registration and target image based on mutual information to carry out registration source images, obtain the transformation relation of two width images, then transformation relation is applied on PET image.Finally two width PET/CT and CLI/CT image are merged based on image pixel, obtained PET/CLI/CT tri-mode image informations, carry out video picture, obtained 68the multi-modality imaging of the animal model for tumour of Ga-AuNRs-RGD labelling.
For a person skilled in the art, can be according to above technical scheme and design, make various corresponding distortion and change, and these all changes and distortion all should be included in the protection domain of the claims in the present invention within.

Claims (5)

  1. Nucleic-Cherenkov based on gold nanorods multiprobe luminous-CT multi-modality imaging method, it is characterized in that, described method comprises the steps:
    Step 1, the preparation of gold nanorods polyfunctional molecule probe:
    Step 1.1, prepares gold nanorods (AuNRs)
    Cetyl trimethyl ammonium bromide (CTAB) and gold chloride are mixed, add sodium borohydride, rapid stirring, obtains crystal seed; Then in the seeded growth liquid that contains silver nitrate, gold chloride, CTAB and ascorbic acid, add gained crystal seed, leave standstill the gold nanorods that obtains specific draw ratio;
    Step 1.2, preparation 68ga-AuNRs-RGD polyfunctional molecule probe
    Adopt small-molecular peptides and the radiosiotope of arginine-glycine-aspartic acid (RGD) tripeptide sequence 68ga carrys out labelling gold nanorods;
    First by difunctional Polyethylene Glycol (OPPS-PEG2K-NHS) bridging agent of excessive different molecular weight respectively with arginine-glycine-aspartic acid (RGD) and 1,4,7,10-tetraazacyclododecanand-1,4,7,10-tetrabasic carboxylic acid (DOTA) mixes, under room temperature, spend the night, form stable bridging agent OPPS-PEG-RGD and OPPS-PEG-DOTA; Then two kinds of bridging agent OPPS-PEG-RGD and OPPS-PEG-DOTA are mixed with the ratio of 2500:2500:1 with described gold nanorods, by the combination of adjacent two thiopyridines bases (OPSS), the stable gold nanorods surface that is connected to of RGD and DOTA;
    Finally, radiotropism isotope in certain proportion 68ga adds the stable gold nanorods coalition being connected of RGD and DOTA, mixes, and reacts 20 minutes, makes 68ga-AuNRs-RGD polyfunctional molecule probe;
    Step 2 is right 68ga-AuNRs-RGD polyfunctional molecule probe carries out Performance Detection:
    Step 2.1, 68ga-AuNRs-RGD polyfunctional molecule probe stability test
    By same concentrations 68ga-AuNRs-RGD polyfunctional molecule probe and gold nanorods are resuspended to respectively in 10% NaCl solution of equivalent, then carry out UVs spectral detection, relatively both stability differences;
    Step 2.2, 68ga-AuNRs-RGD polyfunctional molecule fluorescence probe quantum efficiency and extracorporeal receptor binding analysis
    First, measure 68cherenkov's photo-quantum efficiency of Ga-AuNRs-RGD polyfunctional molecule probe: more different 68same concentration under Ga radioactivity 68cherenkov's flashlight quantum efficiency of Ga-AuNRs-RGD polyfunctional molecule probe, optimization Test parameter, obtains the optimized fluorescence enhancement mode of luminous efficiency 68ga-AuNRs-RGD polyfunctional molecule probe;
    Secondly, measure 68the biology performance of Ga-AuNRs-RGD polyfunctional molecule probe: adopt 68the AuNRs-RGD molecular probe of Ga labelling is as radioligand, and unlabelled AuNRs-RGD molecular probe is non-marked part, analyzes α vβ 3the glioma U87 cell of the expression of receptor positive carries out receptors ligand in conjunction with test; Two kinds of parts are combined with suspension cell, detect their radioactive dosage and luminous efficiency;
    Step 3, tumor bearing nude mice Animal Model:
    In vitro culture glioma U87 cell, in the right shoulder of nude mice place subcutaneous injection of tumor cells several, build Subcutaneous tumor model, after 2-3 week, carry out the operation of subsequent step;
    Step 4, 68the multi-modality imaging of Ga-AuNRs-RGD polyfunctional molecule probe:
    Step 4.1, PET/CT image data acquiring
    Adopt PET/CT imaging system to carry out PET/CT data acquisition, mouse tail vein injection to tumor bearing nude mice 68after Ga-AuNRs-RGD polyfunctional molecule probe, carry out PET/CT scanning at different time points, obtain the PET/CT bimodulus view data of different time points;
    Step 4.2, CLI/CT image data acquiring
    Adopt CLI/CT imaging system to carry out CLI/CT data acquisition, mouse tail vein injection to described tumor bearing nude mice 68after Ga-AuNRs-RGD polyfunctional molecule probe, under airtight dark situation, obtain Cherenkov's luminous signal and the CT view data of multiple spectrum, multiple visual angles toy body surface at different time points;
    Step 4.3, PET/CT, CLI/CT fusing image data
    The PET/CT image that the CLI/CT image drawing in described step 4.2 is drawn in target image and step 4.1 is as source images, through pretreatment such as image denoisings, enhancing, image is cut apart and is carried out feature extraction, mutual information image registration, based on the image co-registration of image pixel, obtain 68ga-AuNRs-RGD polyfunctional molecule probe target to the PET-CLI-CT multi-modality imaging video picture figure of tumor region.
  2. Nucleic-Cherenkov based on gold nanorods multiprobe according to claim 1 luminous-CT multi-modality imaging method, it is characterized in that, in step 1.2 68the molecular weight difference of difunctional Polyethylene Glycol (OPPS-PEG-NHS) bridging agent in the preparation process of Ga-AuNRs-RGD polyfunctional molecule probe: the OPPS-PEG-NHS molecular weight being connected with RGD is 2000, and the OPPS-PEG-NHS molecular weight being connected with DOTA is 5000.
  3. Nucleic-Cherenkov based on gold nanorods multiprobe according to claim 1 luminous-CT multi-modality imaging method, it is characterized in that, in step 3, the operation of subsequent step again after tumor model diameter is greater than 0.5cm after 2-3 week of described Subcutaneous tumor model.
  4. Nucleic-Cherenkov based on gold nanorods multiprobe according to claim 1 luminous-CT multi-modality imaging method, it is characterized in that the α of the RGD target tumor blood vessel on gold nanorods surface in step 4 vβ 3receptor, carries out specific binding to tumor.
  5. Nucleic-Cherenkov based on gold nanorods multiprobe according to claim 1 luminous-CT multi-modality imaging method, it is characterized in that, in step 4, based on the same molecular probe radionuclide in gold nanorods surface 68in the body of Ga, bioprocess carries out respectively optics and radio nuclide imaging.
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