In this study, we developed a highly sensitive dual-mode imaging system using gold nanoparticles （GNPs） conjugated to various fluorophores in solid phantoms. The system consists of fluorescence-lifetime imaging micr...In this study, we developed a highly sensitive dual-mode imaging system using gold nanoparticles （GNPs） conjugated to various fluorophores in solid phantoms. The system consists of fluorescence-lifetime imaging microscopy （FLIM） for surface imaging, diffusion reflection （DR） for deep-tissue imaging （up to 1 cm）, and metal-enhanced fluorescence （MEF）. We detected quenching in the fluorescent intensity （FI） for the conjugation of both gold nanospheres （GNS） and gold nanorods （GNRs） to Fluorescein, which has an excitation peak at a wavelength shorter than the surface plasmon resonance （SPR） of both types of GNPs. Enhanced FI was detected in conjugation to Rhodamine B （RhB） and Sulforhodamine B （SRB）, both with excitation peaks in the SPR regions of the GNPs. The enhanced FI was detected both in solution and in solid phantoms by the FLIM measurements. DR measurements detected the presence of GNRs within the solid phantoms by recording the dropped rates of light scattering in wavelengths corresponding to the absorption spectra of the GNRs. With the inclusion of MEF, this promising dual-mode imaging technique enables efficient and sensitive molecular and functional imaging.展开更多
We successfully conduct the label-free and real-time detection of the interactions between epoxy groups and rabbit IgG and 5＇ CTT CAG GTC ATG AGC CTG AT 3＇ oligonucleotide, and between the hybridization of 5＇ CTT C...We successfully conduct the label-free and real-time detection of the interactions between epoxy groups and rabbit IgG and 5＇ CTT CAG GTC ATG AGC CTG AT 3＇ oligonucleotide, and between the hybridization of 5＇ CTT CAG GTC ATG AGC CTG AT 3＇ and its complementary 3＇ GAA GTC CAC TAC TCG GAC TA 5＇ oligonucleotide, by the oblique-incidence reflectivity difference （OI-RD） method. The dynamic curves of OI-RD signals, corresponding to the kinetic processes of biomolecular combination or hybridization, are acquired. In our case, the combination of epoxy groups with rabbit IgG and 5＇ CTT CAC CTC ATG AGC CTG AT 3＇ oligonucleotide need almost one and a half hours and about two hundred seconds, respectively; and the hybridization of the two oligonucleotides needs about five hundred seconds. The experimental results show that the OI-RD is a promising method for the real-time and label-free detection of biomolecular interactions.展开更多
蛋白质组学研究和药物开发迫切需要高通量、高精度、无标记且实时检测技术。该文提出了一种基于干涉成像的表面等离子共振(SPR)生物分子相互作用检测方法,在入射光的p和s偏振分量之间分别附加90°和-90°位相差,采集对应的干涉...蛋白质组学研究和药物开发迫切需要高通量、高精度、无标记且实时检测技术。该文提出了一种基于干涉成像的表面等离子共振(SPR)生物分子相互作用检测方法,在入射光的p和s偏振分量之间分别附加90°和-90°位相差,采集对应的干涉图像,并利用所采集的2帧图像,计算出反映传感面折射率分布的信号。循环采集能够实时获得生物反应进行时传感表面发生的折射率分布变化。这种方法在克服光源非均匀性影响的同时,还抑制了光强漂移引起的测量误差,达到了高精度。实验结果表明:干涉成像的图像一致性好,系统的折射率分辨率达到了2×10-6RIU(refractive index unit),动态检测区间大于0.005 RIU。在此基础上,检测了兔IgG和羊抗兔IgG的相互作用,结果表明表面等离子体共振干涉成像系统适于高通量检测,完善后有可能成为一种蛋白质组学研究和药物开发的强有力工具。展开更多
文摘In this study, we developed a highly sensitive dual-mode imaging system using gold nanoparticles （GNPs） conjugated to various fluorophores in solid phantoms. The system consists of fluorescence-lifetime imaging microscopy （FLIM） for surface imaging, diffusion reflection （DR） for deep-tissue imaging （up to 1 cm）, and metal-enhanced fluorescence （MEF）. We detected quenching in the fluorescent intensity （FI） for the conjugation of both gold nanospheres （GNS） and gold nanorods （GNRs） to Fluorescein, which has an excitation peak at a wavelength shorter than the surface plasmon resonance （SPR） of both types of GNPs. Enhanced FI was detected in conjugation to Rhodamine B （RhB） and Sulforhodamine B （SRB）, both with excitation peaks in the SPR regions of the GNPs. The enhanced FI was detected both in solution and in solid phantoms by the FLIM measurements. DR measurements detected the presence of GNRs within the solid phantoms by recording the dropped rates of light scattering in wavelengths corresponding to the absorption spectra of the GNRs. With the inclusion of MEF, this promising dual-mode imaging technique enables efficient and sensitive molecular and functional imaging.
基金Project supported by the National Basic Research Program in China(Grant No.2007CB935700)
文摘We successfully conduct the label-free and real-time detection of the interactions between epoxy groups and rabbit IgG and 5＇ CTT CAG GTC ATG AGC CTG AT 3＇ oligonucleotide, and between the hybridization of 5＇ CTT CAG GTC ATG AGC CTG AT 3＇ and its complementary 3＇ GAA GTC CAC TAC TCG GAC TA 5＇ oligonucleotide, by the oblique-incidence reflectivity difference （OI-RD） method. The dynamic curves of OI-RD signals, corresponding to the kinetic processes of biomolecular combination or hybridization, are acquired. In our case, the combination of epoxy groups with rabbit IgG and 5＇ CTT CAC CTC ATG AGC CTG AT 3＇ oligonucleotide need almost one and a half hours and about two hundred seconds, respectively; and the hybridization of the two oligonucleotides needs about five hundred seconds. The experimental results show that the OI-RD is a promising method for the real-time and label-free detection of biomolecular interactions.
文摘蛋白质组学研究和药物开发迫切需要高通量、高精度、无标记且实时检测技术。该文提出了一种基于干涉成像的表面等离子共振(SPR)生物分子相互作用检测方法,在入射光的p和s偏振分量之间分别附加90°和-90°位相差,采集对应的干涉图像,并利用所采集的2帧图像,计算出反映传感面折射率分布的信号。循环采集能够实时获得生物反应进行时传感表面发生的折射率分布变化。这种方法在克服光源非均匀性影响的同时,还抑制了光强漂移引起的测量误差,达到了高精度。实验结果表明:干涉成像的图像一致性好,系统的折射率分辨率达到了2×10-6RIU(refractive index unit),动态检测区间大于0.005 RIU。在此基础上,检测了兔IgG和羊抗兔IgG的相互作用,结果表明表面等离子体共振干涉成像系统适于高通量检测,完善后有可能成为一种蛋白质组学研究和药物开发的强有力工具。
