用于疾病诊断的Gd~Ⅲ/量子点多模态成像探针的构建

结合核磁共振成像(MRI)和荧光成像技术,以钆离子、近红外低毒量子点、二氧化硅和聚丙烯酸(PAA)等为原料,采用一系列纳米载体自组装技术,构建出MRI弛豫率/荧光效率高和生物相容性好的GdⅢ/量子点多模态纳米探针.结果表明,与未螯合GdⅢ的量子点纳米探针相比,GdⅢ/量子点多模态纳米探针具有更高的弛豫率;t1-加权MRI成像也证实了GdⅢ/量子点多模态纳米探针具有很好的阳性造影功效.     

Gd3+/Yb3+掺杂ZnO量子点作为双模式磁共振/CT成像探针

基于结合磁共振成像(MRI)的高空间分辨率和CT成像的深穿透能力设计思想,在乙醇溶液中水解醋酸锌、醋酸钆和醋酸镱,制备了油酸稳定的Gd3+/Yb3+掺杂ZnO量子点(ZnO∶Gd/Yb),并对其表面进行了氨基修饰。研究了ZnO∶Gd/Yb量子点的弛豫性能、X射线吸收性能、细胞毒性及体外MRI和CT成像。当Zn2+,Gd3+,Yb3+的摩尔比为1.0∶0.12∶0.20时,ZnO∶Gd/Yb量子点展现了最高的弛豫效率6.06 mmol/(L.s)对X-射线的吸收能力也显著高于临床CT造影剂碘比醇。体外MRI和CT成像实验表明,当Gd3+的浓度为1.5 mmol/L时,T1加权MRI信号明显增强,当Yb3+的浓度为5 g/L时,可呈现清晰的CT图像。细胞毒性实验表明,ZnO∶Gd/Yb量子点的浓度低于1.5 mmol/L(Gd3+)时,量子点的毒性相对较低。     

钆掺杂碳量子点的制备及其双模态成像研究进展

荧光-磁共振成像(MRI)双模态分子探针突破了单一分子影像探针在灵敏度以及软组织分辨率等方面的局限性,在诊断和治疗等医学领域具有良好的应用潜力。钆掺杂碳量子点(Gd-CDs)是一种典型的荧光-MRI双模态分子探针,因其优异的荧光性能、良好的核磁共振成像能力和生物相容性等优点,在肿瘤的靶向性成像和监控诊疗方面具有广泛的应用前景。本文综述了目前Gd-CDs的制备、性能及其在双模态成像应用的研究现状,并提出目前存在的问题和前景展望。     

靶向髓过氧化酶的双模态探针分子的合成与表征

5-羟色胺(5-HT)对动脉粥样硬化斑块中巨噬细胞分泌的髓过氧化酶(MPO)具有靶向功能,通过酯化反应将5-HT接枝到两亲型聚合物聚衣康酸接枝聚乙二醇接枝十二胺(PIA-g-PEG-g-DDA)上,制备了具有靶向功能的两亲型聚合物(5-HT-g-PIA-g-PEG-g-DDA).采用配体交换法,将超顺磁性氧化铁纳米粒子(SPION)包覆在该聚合物上制备具有磁性的纳米粒子(5-HT-g-PIA-g-PEG-g-DDA@SPION),通过静电力结合使其与碳点(CDs)结合,制备具有荧光成像和核磁造影成像的双模态探针的纳米粒子.采用动态光散射(DLS)及透射电子显微镜(TEM)分析了其粒径和形貌.采用荧光光谱仪与振动样品磁强计(VSM)测试证明该纳米粒子具备双模态成像功能.共聚焦成像(CLSM)的测试结果表明,该纳米试剂对动脉粥样硬化斑块中的巨噬细胞具有靶向性,MTT法的测试结果表明该探针分子具有良好的生物相容性.该双模态探针具有靶向性好、分辨率高及使用便捷的特点,是一种性能优异的双模态检测纳米试剂.     

CdTe/Mn3 O4/SiO2荧光/磁共振双模成像纳米球的制备与表征

成功地制备了CdTe/Mn_3O_4/SiO_2核壳结构的荧光/磁共振成像双功能纳米球,并用透射电镜(TEM)、能谱分析(EDXA)、磁共振成像(MRI)、红外、荧光光谱等对其结构、磁共振成像和发光性能进行了表征。TEM照片显示所合成的纳米球具有明显的球形核壳结构。EDXA分析显示所制备的CdTe/Mn_3O_4/SiO_2纳米球表面只检测到Si和O元素,证明CdTe量子点和Mn_3O_4纳米立方体被成功地包被于二氧化硅纳米球之内。荧光发射光谱显示相对于CdTe量子点,CdTe/Mn_3O_4/SiO_2纳米球荧光发射光谱虽然发生了一定的蓝移,但是仍具有良好的荧光性能。MRI分析可知CdTe/Mn_3O_4/SiO_2纳米球的弛豫参数(r_1)为3.88 s~(-1)(mg·L~(-1))~(-1),说明所合成的CdTe/Mn_3O_4/SiO_2纳米球可用于T_1-加权磁共振成像。细胞毒性实验表明,当CdTe/Mn_3O_4/SiO_2溶液浓度达到300μg·mL~(-1)时,细胞活力仍可达到90%以上,表明此浓度对细胞的毒性作用较弱。     

基于银纳米粒子-CdTe量子点复合物荧光探针检测水样和血清中L-半胱氨酸

利用无荧光的碳量子点-银纳米颗粒(CQDs-AgNPs)中Ag+与CdTeQDs表面S2-的强亲和作用,通过S-Ag键形成弱荧光的CQDs-AgNPs/CdTe QDs纳米复合物探针.L-半胱氨酸(L-Cys)中-S-与复合物探针中Ag+竞争络合,可使CdTe QDs的荧光恢复,基于L-Cys对复合物探针的荧光恢复作用...     

NaYF4∶Yb3+,Er3+@NaGdF4@TaOx多模态纳米探针的合成及其在生物成像中的应用

采用溶剂热法在稀土掺杂NaYF4∶Yb3+,Er3+上转换纳米粒子上包覆一层NaGdF4,再利用反相微乳液法在NaGdF4上覆盖一层TaOx,从而合成NaYF4∶Yb3+,Er3+@NaGdF4@TaOx核壳壳结构的纳米探针。使用透射电镜(TEM)、X射线衍射(XRD)及X射线能量色散谱分析(EDS)对NaYF4∶Yb3+,Er3+@NaGdF4@TaOx纳米探针的结构和组成进行表征。并通过荧光光谱、磁滞回线以及电子计算机X射线横断扫描(CT)造影成像等方法对其性能进行了表征,证明该纳米探针具有良好的光学、磁学和CT造影特性。将NaYF4∶Yb3+,Er3+@NaG-dF4@TaOx纳米探针应用于小鼠活体成像实验,结果表明,这种纳米探针对小鼠肿瘤部位的磁共振成像(MRI)和CT信号均有较好的增强效果,表明其在多模态造影成像方面有潜在的应用前景。     

基于量子点-甲酚紫FRET比率探针的活细胞硫化氢成像

硫化氢(H_2S)作为一种重要的气体信号分子,其细胞水平异常与多种疾病的发生发展密切相关。本研究针对现存荧光探针的胞内非特异性响应问题,发展了一种基于量子点-甲酚紫的荧光能量共振转移(F9rster resonance energy transfer,FRET)探针,利用比率荧光分析,实现活细胞内H_2S的灵敏成像。通过一步超声乳化法制备量子点纳米球,再结合叠氮甲酚紫(CV-N_3)制得无机-有机杂化QDS-N3比率探针。此纳米探针尺寸均一,粒径约为120 nm,酸性稳定性高,且无细胞毒性。H2S可将CV-N_3上的N3还原成NH2,生成的氨基甲酚紫(CV-NH_2)作为FRET-受体,吸收量子点纳米球的橙色荧光,产生红光信号。通过两个波长的荧光强度的比率分析可有效消除光源强度波动和细胞内复杂环境等的干扰,最终实现了活细胞内H2S成像。     

GdPO4:Eu3+和GdBO3:Eu3+中"Gd-Eu"间的能量传递

利用同步辐射光源(德国HASYLAB实验室的SUPERLUMI实验站)对高温固相法制备的GdPO4 Eu3+和GdBO3Eu3+的发射谱与激发谱进行了研究, 并从量子剪裁的角度对"Gd3+-Eu3+"之间的能量传递作了讨论. 在两种样品监测Eu3+红光发射的激发谱上, 都显著观察到了对应Gd3+离子4f-4f跃迁的谱线, 说明从基质中Gd3+离子向掺杂的Eu3+离子之间存在非常有效的能量传递, 这有利于量子剪裁的发生. 在激发谱上还观察到了对应Eu3+-O2-电荷迁移态的宽激发带(180～270 nm), 而对应Gd3+离子8S7/2→6GJ跃迁的谱线(196 nm, 203 nm)叠加在这个宽带之上, 微弱可辨, 这不利于Gd3+-Eu3+间交叉弛豫的能量传递过程, 从而不利于量子剪裁的发生. 结合Eu3+-O2-电荷迁移态位置变化的规律, 提出了如何避开其干扰以增强量子剪裁效应的材料设计方案.     

荧光/MRI双模态靶向成像诊疗试剂的制备

在聚乙二醇二胺(NH_2-PEG-NH_2)修饰的石墨烯量子点(GODs)表面以酰胺键偶联二乙基三胺五乙酸(DTPA)分子,之后将Gd~(3+)离子与其进行配合,得到了GODs-Gd(DTPA)复合纳米粒子,然后再通过酰胺键在GODs-Gd(DTPA)的表面修饰叶酸(FA)靶分子,最后进一步将阿霉素(DOX)通过π-π堆垛吸附在造影剂的表面,制备了FA/GODs-Gd(DTPA)/DOX荧光/MRI双模态靶向肺癌细胞成像诊疗试剂,通过透射电子显微镜、紫外可见吸收光谱、荧光光谱和激光共聚焦扫描显微镜等手段表征了其形貌、发光性能和靶向成像性能。MRI、激光共聚焦扫描显微镜和MTT等结果表明,相对于正常的HLF细胞,所制备的FA/GODs-Gd(DTPA)/DOX纳米粒子能够靶向检测FA受体高表达的肺癌H460细胞,并具有明显的抗肿瘤活性。     

Electrostatically assembled biocompatible polymer nanoparticles for MR/optical dual-modality imaging nanoprobes

Biocompatible dual-modality imaging nanoprobes were synthesized by the electrostatic assembly of poly(γ-glutamic acid)[Gd-DTPA] and chitosan[IRDye800] and applied for the imaging of immune cells (phagocytic) and cancer cells (non-phagocytic).     

Solute-solvent halogen bonding during adsorption on silver nanostructures

Multimodal imaging technique is an alternative approach to improve sensitivity of early cancer diagnosis. In this study, highly fluorescent and strong X-ray absorption coefficient gold nanoclusters (Au NCs) are synthesized as dual-modality imaging contrast agents (CAs) for fluorescent and X-ray dual-modality imaging. The experimental results show that the as-prepared Au NCs are well constructed with ultrasmall sizes, reliable fluorescent emission, high computed tomography (CT) value and fine biocompatibility. In vivo imaging results indicate that the obtained Au NCs are capable of fluorescent and X-ray enhanced imaging.     

Multifunctional nanoprobe for cancer cell targeting and simultaneous fluorescence/magnetic resonance imaging

Multifunctional nanoprobes with distinctive magnetic and fluorescent properties are highly useful in accurate and early cancer diagnosis. In this study, nanoparticles of Fe₃O₄ core with fluorescent SiO₂ shell (MFS) are synthesized by a facile improved Stöber method. These nanoparticles owning a significant core-shell structure exhibit good dispersion, stable fluorescence, low cytotoxicity and excellent biocompatibility. TLS11a aptamer (Apt1), a specific membrane protein for human liver cancer cells which could be internalized into cells, is conjugated to the MFS nanoparticles through the formation of amide bond working as a target-specific moiety. The attached TLS11a aptamers on nanoparticles are very stable and can't be hydrolyzed by DNA hydrolytic enzyme in vivo. Both fluorescence and magnetic resonance imaging show significant uptake of aptamer conjugated nanoprobe by HepG2 cells compared to 4T1, SGC-7901 and MCF-7 cells. In addition, with the increasing concentration of the nanoprobe, T₂-weighted MRI images of the as-treated HepG2 cells are significantly negatively enhanced, indicating that a high magnetic field gradient is generated by MFS-Apt1 which has been specifically captured by HepG2 cells. The relaxivity of nanoprobe is calculated to be 11.5 mg⁻¹s⁻¹. The MR imaging of tumor-bearing nude mouse is also confirmed. The proposed multifunctional nanoprobe with the size of sub-100 nm has the potential to provide real-time imaging in early liver cancer cell diagnosis.     

点击功能化的叶酸受体靶向荧光纳米探针用于肿瘤细胞成像

采用点击化学偶联法对荧光二氧化硅纳米粒子表面进行叶酸功能化修饰,构建了一种叶酸受体靶向的荧光纳米探针,并成功用于肿瘤细胞的成像研究.首先通过St?ber法制备包裹钌联吡啶的荧光二氧化硅纳米粒子(RSiNPs),然后利用叠氮化硅烷偶联剂(Az-PTES)的水解反应在其表面引入叠氮基团,最后通过点击化学反应将炔丙基叶酸衍生物偶联到粒子表面.利用红外光谱对其偶联前后的叠氮基特征峰(2105 cm-1)进行表征,证实了叶酸功能化的荧光纳米探针(RSiNPs-Folate)已被成功制备.在生理pH条件下,以458 nm为激发波长,RSiNPs-Folate在601 nm处发射较强的红色荧光,且光稳定性较好.细胞成像结果表明,这种叶酸受体靶向的荧光纳米探针能够有效地标记叶酸受体呈阳性的人宫颈癌细胞(HeLa),而叶酸受体呈阴性的人肺癌细胞(A549)未观察到明显的荧光.叶酸竞争性结合实验证明了这种叶酸受体介导的肿瘤细胞成像机制.此探针能够实现混合细胞体系中HeLa细胞的选择性识别与荧光成像.与酰胺化反应偶联叶酸相比,这种点击功能化的纳米探针的合成方法简单、反应条件温和、产率高,可用于不同肿瘤细胞的荧光标记与成像.     

Red carbon dots as label-free two-photon fluorescent nanoprobes for imaging of formaldehyde in living cells and zebrafishes

Direct,in situ selective detection of intracellular formaldehyde(FA)is of great significance for understanding its function in FA-related diseases.Herein,red carbon dots(RCD)are reported as label-free two-photon fluorescent nanoprobes for detecting and imaging of FA.Upon addition of FA,the-NH2 groups of RCD could quickly and specially react with aldehydes to form Schiff base and then the strong fluorescence of RCD with blue-shift emission is recovery due to the destruction of the hydrogen bond interaction between RCD and water.In addition,the nanoprobes exhibit outsta nding photo stability,rapid response(<1 min),high sensitivity(~9.9μmol/L)and excellent selectivity toward FA over other aldehyde group compounds.Notably,owing to the good cell-membrane permeability and biocompatibility,as well as the large two-photon absorption cross-section,the as-prepared RCD can be used as label-free nanoprobes for selectively detecting and imaging FA in living cells and zebrafishes through one-photon and two-photon excitation.Moreover,RCD could stain the tissue of zebrafishes at depths interval of up to 240μm under two-photon excitation.This research implied that RCD are promising tools for directly and in situ imaging FA in vivo,thus providing critical insights into FA-related pathophysiological processes.     

Quantum dot capped magnetite nanorings as high performance nanoprobe for multiphoton fluorescence and magnetic resonance imaging

In the present study, quantum dot (QD) capped magnetite nanorings (NRs) with a high luminescence and magnetic vortex core have been successfully developed as a new class of magnetic-fluorescent nanoprobe. Through electrostatic interaction, cationic polyethylenimine (PEI) capped QD have been firmly graft into negatively charged magnetite NRs modified with citric acid on the surface. The obtained biocompatible multicolor QD capped magnetite NRs exhibit a much stronger magnetic resonance (MR) T2* effect where the r2* relaxivity and r2*/r1 ratio are 4 times and 110 times respectively larger than those of a commercial superparamagnetic iron oxide. The multiphoton fluorescence imaging and cell uptake of QD capped magnetite NRs are also demonstrated using MGH bladder cancer cells. In particular, these QD capped magnetite NRs can escape from endosomes and be released into the cytoplasm. The obtained results from these exploratory experiments suggest that the cell-penetrating QD capped magnetite NRs could be an excellent dual-modality nanoprobe for intracellular imaging and therapeutic applications. This work has shown great potential of the magnetic vortex core based multifunctional nanoparticle as a high performance nanoprobe for biomedical applications.     

Synthesis and characterization of fluorescent magneto polymeric nanoparticles (FMPNs) for bimodal imaging probes

Novel bifunctional fluorescent magneto polymeric nanoprobes (FMPNs) were synthesized to provide simultaneous diagnostic information via magnetic resonance imaging (MRI) and optical imaging. FMPNs consist of ultra-sensitive magnetic nanocrystals that function as MR probes combined with Nile Red, which functions as a fluorescent probe. FMPNs were encapsulated by a nano-emulsion method in polyvinyl alcohol (PVA, 87–89% hydrolyzed) through a matrix of polymethyl methacrylate (PMMA). FMPNs exhibited excellent colloidal stability and monodispersity. The production of MR and optical images demonstrated that FMPNs have potential as dual-mode imaging agents.     

Folate-targeted optical and magnetic resonance dualmodality PCL-<Emphasis Type="Italic">b</Emphasis>-PEG micelles for tumor imaging

A biodegradable tumor targeting nano-probe based on poly（ε-caprolactone）-b-poly（ethylene glycol）block copolymer（PCL-b-PEG）micelle functionalized with a magnetic resonance imaging（MRI）contrast agent diethylenetriaminepentaacetic acid-gadolinium(DTPA-Gd³⁺)on the shell and a near-infrared（NIR）dye in the core for magnetic resonance and optical dual-modality imaging was prepared.The longitudinal relaxivity（r₁）of the PCL-b-PEG-DTPA -Gd³⁺micelle was 13.4（mmol/L）^-1s^-1,three folds of that of DTPA-Gd³⁺,and higher than that of many polymeric contrast agents with similar structures.The in vivo optical imaging of a nude mouse bearing xenografted breast tumor showed that the dual-modality micelle preferentially accumulated in the tumor via the folic acid-mediated active targeting and the passive accumulation by the enhanced permeability and retention（EPR）effect.The results indicated that the dualmodality micelle is a promising nano-probe for cancer detection and diagnosis.     

荧光纳米探针用于细胞器pH检测的研究进展

pH稳态对于维持活细胞细胞器的正常功能具有重要作用. 细胞器内pH稳态被打破会导致细胞器功能的紊乱, 进而引发癌症、 神经退行性疾病等相关疾病. 因此, 在活细胞水平上定量测定pH并对其波动进行实时监测对于理解相关疾病的发生机制非常重要. 基于非侵入、 高时空分辨率成像的优势, 荧光探针非常适合用于活细胞内pH的检测. 本综述总结了近些年利用不同种类荧光纳米探针对不同细胞器进行pH成像的研究工作, 并对荧光纳米探针应用面临的机遇与挑战进行了展望.