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71.
Plaque angiogenesis and plaque hemorrhage are major players in the destabilization and rupture of atherosclerotic lesions. As these are dynamic processes, imaging of plaque angiogenesis, especially the integrity or leakiness of angiogenic vessels, can be an extremely useful tool in the studies on atherosclerosis pathophysiology. Visualizing plaque microvessels in 3D would enable us to study the architecture and permeability of adventitial and intimal plaque microvessels in advanced atherosclerotic lesions. We hypothesized that a comparison of the vascular permeability between healthy continuous and fenestrated as well as diseased leaky microvessels, would allow us to evaluate plaque microvessel leakiness. We developed and validated a two photon intravital microscopy (2P-IVM) method to assess the leakiness of plaque microvessels in murine atherosclerosis-prone ApoE3*Leiden vein grafts based on the quantification of fluorescent-dextrans extravasation in real-time. We describe a novel 2P-IVM set up to study vessels in the neck region of living mice. We show that microvessels in vein graft lesions are in their pathological state more permeable in comparison with healthy continuous and fenestrated microvessels. This 2P-IVM method is a promising approach to assess plaque angiogenesis and leakiness. Moreover, this method is an important advancement to validate therapeutic angiogenic interventions in preclinical atherosclerosis models.  相似文献   
72.
针对所研制中的一台多轴双光子加工系统,通过齐次坐标变换建立了激光束焦斑中心与试件期望位置之间的综合误差模型。假定对各个运动轴误差及光路系统的误差服从非平稳概率分布,考察了各个运动轴误差及光路系统误差对综合误差的影响。为主动补偿多轴双光子聚合加工系统的综合误差、提升多轴双光子聚合加工系统的工作精度提供了依据。  相似文献   
73.
Multiphoton micro-spectroscopy, employing diffraction optics and electron-multiplying CCD (EMCCD) cameras, is a suitable method for determining protein complex stoichiometry, quaternary structure, and spatial distribution in living cells using Förster resonance energy transfer (FRET) imaging. The method provides highly resolved spectra of molecules or molecular complexes at each image pixel, and it does so on a timescale shorter than that of molecular diffusion, which scrambles the spectral information. Acquisition of an entire spectrally resolved image, however, is slower than that of broad-bandwidth microscopes because it takes longer times to collect the same number of photons at each emission wavelength as in a broad bandwidth. Here, we demonstrate an optical micro-spectroscopic scheme that employs a laser beam shaped into a line to excite in parallel multiple sample voxels. The method presents dramatically increased sensitivity and/or acquisition speed and, at the same time, has excellent spatial and spectral resolution, similar to point-scan configurations. When applied to FRET imaging using an oligomeric FRET construct expressed in living cells and consisting of a FRET acceptor linked to three donors, the technique based on line-shaped excitation provides higher accuracy compared to the point-scan approach, and it reduces artifacts caused by photobleaching and other undesired photophysical effects.  相似文献   
74.
建立一台基于同步扫描相机的双光子激发荧光寿命显微测量系统,同步扫描相机的重复工作频率为76 MHz,利用钛宝石飞秒激光器作为光源,通过可调延时器和标准具对扫描相机的时间分辨率、扫描速度以及非线性等进行标定.该系统的时间分辨率为9 ps,非线性小于4%,量程为2.8 ns.测量了荧光染料Rose Bengal(RsB)的荧光衰减曲线,通过最小二乘法对荧光的衰减曲线进行拟合,得到RsB的荧光寿命为763 ps,与标准荧光染料对比一致.  相似文献   
75.
双光子吸收(two-photon absorption)是介质通过中间虚态同时吸收两个光子从基态跃迁至激发态的过程;其跃迁量子特征具有高度空间分辨率和良好介质穿透性,因而在三维微纳加工制作、三维光信息存储和三维荧光显微成像等高科技领域具有应用价值,成为当前有机光电子领域热点课题。该文介绍本课题组十年来在双光子科学领域中开展的工作与相关成果。  相似文献   
76.
The relationships between two-photon absorption (TPA) effect and excited state intramolecular charge transfer (ICT) of p-substituted stilbene-type pyridinium salts have been investigated. Two-photon absorption cross section (δTPA) for all the compounds has been determined by open aperture Z-scanning technique, while two-photon pumped (TPP) up-converted lasing efficiencies (η) and optical limiting behavior have been measured by two-channel energy-meter, pumped by a mode locked Nd: YAG ps laser. Streak camera was used to record two-photon induced fluorescence and the fluorescence lifetimes. The experimental and calculated results have shown that the excited state larger in-tra-molecular charge transfer, in the excited state characterized by the charge density difference between the ground state ( S0) and the first excited state ( S1), has contributed to enhancing TPA cross section and may result in better optical limiting effect. And high TPP up-converted lasing efficiency is related to not only the δTP  相似文献   
77.
Neurotransmitter uncaging, especially that of glutamate, has been used to study synaptic function for over 30 years. One limitation of caged glutamate probes is the blockade of γ‐aminobutyric acid (GABA)‐A receptor function. This problem comes to the fore when the probes are applied at the high concentrations required for effective two‐photon photolysis. To mitigate such problems one could improve the photochemical properties of caging chromophores and/or remove receptor blockade. We show that addition of a dicarboxylate unit to the widely used 4‐methoxy‐7‐nitroindolinyl‐Glu (MNI‐Glu) system reduced the off‐target effects by about 50–70 %. When the same strategy was applied to an electron‐rich 2‐(p‐Phenyl‐o‐nitrophenyl)propyl (PNPP) caging group, the pharmacological improvements were not as significant as in the MNI case. Finally, we used very extensive biological testing of the PNPP‐caged Glu (more than 250 uncaging currents at single dendritic spines) to show that nitro‐biphenyl caging chromophores have two‐photon uncaging efficacies similar to that of MNI‐Glu.  相似文献   
78.
A photoresist system for 3D two-photon microprinting is presented, which enables the printing of inherently nanoporous structures with mean pore sizes around 50 nm by means of self-organization on the nanoscale. A phase separation between polymerizable and chemically inert photoresist components leads to the formation of 3D co-continuous structures. Subsequent washing-out of the unpolymerized phase reveals the porous polymer structures. To characterize the volume properties of the printed structures, scanning electron microscopy images are recorded from ultramicrotome sections. In addition, the light-scattering properties of the 3D-printed material are analyzed. By adjusting the printing parameters, the porosity can be controlled during 3D printing. As an application example, a functioning miniaturized Ulbricht light-collection sphere is 3D printed and tested.  相似文献   
79.
Developing small-scale, lightweight, and flexible devices with integrated microactuators is one of the critical challenges in wearable haptic devices, soft robotics, and microrobotics. In this study, a novel fabrication process that leverages the benefits of 3D printing with two-photon polymerization and flexible printed circuit boards (FPCBs) is presented. This method enables flexible microsystems with 3D-printed electrostatic microactuators, which are demonstrated in a flexible integrated micromirror array and a legged microrobot with a mass of 4 mg. 3D electrostatic actuators on FPCBs are robust enough to actuate the micromirrors while the device is deformed, and they are easily integrated with off-the-shelf electronics. The crawling robot is one of the lightest legged microrobots actuated without external fields, and the legs actuated with 3D electrostatic actuators enable a locomotion speed of 0.27 body length per second. The proposed fabrication framework opens up a pathway toward a variety of highly integrated flexible microsystems.  相似文献   
80.
Two-photon polymerization (TPP) is widely used to create 3D micro- and nanoscale scaffolds for biological and mechanobiological studies, which often require the mechanical characterization of the TPP fabricated structures. To satisfy physiological requirements, most of the mechanical characterizations need to be conducted in liquid. However, previous characterizations of TPP fabricated structures are all conducted in air due to the limitation of conventional micro- and nanoscale mechanical testing methods. In this study, a new experimental method is reported for testing the mechanical properties of TPP-printed microfibers in liquid. The experiments show that the mechanical behaviors of the microfibers tested in liquid are significantly different from those tested in air. By controlling the TPP writing parameters, the mechanical properties of the microfibers can be tailored over a wide range to meet a variety of mechanobiology applications. In addition, it is found that, in water, the plasticly deformed microfibers can return to their predeformed shape after tensile strain is released. The shape recovery time is dependent on the size of microfibers. The experimental method represents a significant advancement in mechanical testing of TPP fabricated structures and may help release the full potential of TPP fabricated 3D tissue scaffolds for mechanobiological studies.  相似文献   
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