84 resultados para total internal reflection fluorescence microscopy
Resumo:
Whispering gallery modes (WGMs) in microcavities possess ultra-high cavity Q factor. Such microcavity are easy to be fabricated, so WGMs have attracted much attention in the area of photonics and integrated photonic circuits. It is well known that the effect of total internal reflection restricts the size of this mirocavity. Such drawback goes against the integration of photon. However, the photonic crystal microcavities (PCMC) make a breakthrough recently. The WGMs in the PCMC are possible to gain both ultra-high Q and ultra-small mode volume. In this paper, the property of the mode in photonic crystal ring cavity is analyzed by FDTD and PWE. By modifying the airholes in the corners of the ring cavity, we can obtain the WGM. Also the Q factor of WGM in photonic crystal ring cavity is calculated. This favors the design of the photonic crystal microcavity components.
Resumo:
SOI (Silicon on Insulator) based photonic devices, including stimulated emission from Si diode, RCE (Resonant Cavity Enhanced) photodiode with quantum structure, MOS (Metal Oxide Semiconductor) optical modulator with high frequency, SOI optical matrix switch and wavelength tunable filter are reviewed in the paper. The emphasis will be played on our recent results of SOI-based thermo-optic waveguide matrix switch with low insertion loss and fast response. A folding re-arrangeable non-blocking 4x4 matrix switch with total internal reflection (TIR) mirrors and a first blocking 16 x 16 matrix were fabricated on SOI wafer. The extinction ratio and the crosstalk are better. The insertion loss and the polarization dependent loss (PDL) at 1.55 mu m increase slightly with longer device length and more bend and intersecting waveguides. The insertion losses are expected to decrease 2-3 dB when anti-reflection films are added in the ends of the devices. The rise and fall times of the devices are 2.1 mu s and 2.3 mu s, respectively.
Resumo:
The electrochemical behavior of horseradish peroxidase (HRP) in the dimyristoyl phosphatidylcholine (DMPC) bilayer on the glassy carbon (GC) electrode was studied by cyclic voltammetry. The direct electron transfer of HRP was observed in the DMPC bilayer. Only a small cathodic peak was observed for HRP on the bare GC electrode. The electron transfer of HRP in the DMPC membrane is facilitated by DMPC membrane. UV-Vis and circular dichroism (CD) spectroscopy were used to study the interaction between HRP and DMPC membrane. On binding to the DMPC membrane the secondary structure of HRP remains unchanged while there is a substantial change in the conformation of the heme active site. Tapping mode atomic force microscopy (AFM) was first applied for the investigation on the structure of HRP adsorbed on supported phospholipid bilayer on the mica and on the bare mica. HRP molecules adsorb and aggregate on the mica without DMPC bilayer. The aggregation indicates an attractive interaction among the adsorbed molecules. The molecules are randomly distributed in the DMPC bilayer. The adsorption of HRP in the DMPC bilayer changes drastically the domains and defects in the DMPC bilayer due to a strong interaction between HRP and DMPC films.
Resumo:
We theoretically demonstrate that enhanced penetration depth in three-dimensional multiphoton microscopy can be achieved using concentric two-color two-photon (C2C2P) fluorescence excitation in which the two excitation beams are separated in space before reaching their common focal spot. Monte Carlo simulation shows that, in comparison with the one-color two-photon excitation scheme, the C2C2P fluorescence microscopy provides a significantly greater penetration depth for imaging into a highly scattering medium. (C) 2008 Optical Society of America.
Resumo:
The fluorescence emission from indole resulting from two-color two-photon (2C2P) excitation with 400 and 800 nm wavelengths is observed, using the second harmonic and fundamental wavelength of a 800 nm 40 fs pulsed Ti:Sapphire femtosecond (fs) regenerative amplifier operating at a repetition rate of 1 kHz. By delaying one fs laser pulse relative to the other, the cross correlation of fluorescence is observed, which indicates the generation of 2C2P fluorescence signal in the experiment. The strongest 2C2P fluorescence emission characterized by the peak of cross correlation curve suggests optimal temporal overlap of the two fs laser pulses. The 2C2P fluorescence signal is linearly dependent on the total excitation intensity. The fluorescence signals with 400 nm and 800 nm irradiation alone are also demonstrated and discussed in this paper. (C) 2008 Elsevier B.V. All rights reserved.
Resumo:
The structure of the inhibition patterns is important to the stimulated emission depletion (STED) microscopy. Usually, Laguerre-Gaussian (LG) beam and the central zero-intensity patterns created by inserting phase masks in Gaussian beams are used as the erase beam in STED microscopy. Aberration is generated when focusing beams through an interface between the media of the mismatched refractive indices. By use of the vectorial integral, the effects of such aberration on the shape of depletion patterns and the size of fluorescence emission spot in the STED microscopy are studied. Results are presented as a comparison between the aberration-free case and the aberrated cases. (C) 2009 Optical Society of America
Resumo:
Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to beta(2) subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively. (c) 2006 Elsevier Inc. All rights reserved.
Resumo:
We investigate the large negative lateral displacements of TE polarized light beams reflected from or transmitted through an active slab surrounded by transparent medium. The large negative displacements can be achieved when the incidence angle of the beam is less than but close to the critical angle for total reflection. It is also shown that both the reflectivity and transmissivity of the beam that correspond to the large negative displacements can be enhanced by active medium. These phenomena may lead to convenient measurements and interesting applications in optical devices. (c) 2006 Elsevier B.V. All rights reserved.
Resumo:
We present a multifunctional darkfield microscopy using an axicon. It combines the functions of a darkfield microscope, fluorescence microscope, and microspectrophotometer in one platform. The advantage of the system over conventional darkfield microscopy includes the high transmittance of the illuminating flux, the high contrast of the image, and the convenient toggle between darkfield and brightfield microscopy. Examples of dark, bright, and fluorescent micrographs as well as concerned spectra of microsized specimens implemented in this apparatus are demonstrated. (C) 2008 Society of Photo-Optical Instrumentation Engineers.
Resumo:
Atomic force microscopy (AFM) and lateral force microscopy (LFM) were used simultaneously to analyze a model membrane bilayer structure consisting of a phospholipid outer monolayer deposited onto organosilane-derivatized mica surfaces, which were constructed by using painting and self-assembly methods. The phospholipid used as outer monolayer was dimyristoylphosphatidylcholine (DMPC). The hydrocarbon-covered substrate that formed the inner half bilayer was composed of a self-assembly monolayer (SAM) of octadecyltrichloroorganosilane (OTS) on mica. SAMs of DMPC were formed by exposing hydrophobic mica to a solution of DMPC in decane/isobutanol and subsequently immersing into pure water. AFM images of samples immersed in solution for varying exposure times showed that before forming a complete monolayer the molecules aggregated into dense islands (2.2-2.6 nm high) on the surface. The islands had a compact and rounded morphology. LFM, coupled with topographic data obtained with the atomic force mode, had made possible the distinction between DMPC and OTS. The rate constant of DMPC growth was calculated. This is the first systematic study of the SAM formation of DMPC by AFM and LFM imaging. It reveals more direct information about the film morphology than previous studies with conventional surface analytical techniques such as infrared spectroscopy, X-ray, or fluorescence microscopy.
Resumo:
受激发射损耗荧光显微镜利用荧光饱和和激发态荧光受激损耗的非线性关系,通过限制损耗区域,可突破远场光学显微术的衍射极限分辨力并实现三维成像。基于对粒子速率方程组的修正,建立了描述荧光团各能级粒子数概率时间特性的模型,并定义了时间平均损耗效率判据。采用高斯函数模拟两束入射激光脉冲通过对模型的数值计算,模拟了激发脉冲的S1ED激光脉冲的光强、脉冲宽度以及两束光的延迟时间等参量与损耗效率之间的关系,并获得了各参量的最佳值,优化r损耗效率,为提高系统分辨力提供了有效的途径。
Resumo:
长期以来,远场光学荧光显微镜凭借其非接触、无损伤、可探测样品内部等优点,一直是生命科学中最常用的观测工具。但由于衍射极限的存在,使传统的宽场光学显微镜横向和纵向的分辨率分别仅约为230 nm和1000 nm。为了揭示细胞内分子尺度的动态和结构特征,提高光学显微镜分辨率成为生命科学发展的迫切要求,在远场荧光显微镜的基础上,科学家们已经发展出许多实用的提高分辨率甚至超越分辨率极限的成像技术。例如,采用横向结构光照明提高横向分辨率到约100 nm,利用纵向驻波干涉效应将纵向分辨率提高5~10倍。然而,直到在光学荧光显微镜中引入非线性效应后,衍射极限才被真正突破,如受激荧光损耗显微镜利用非线性效应实现了30~50 nm的三维分辨率。另外应用荧光分子之间能量转移共振原理以及单荧光分子定位技术也可以突破衍射极限,甚至可以将分子定位精度提高到几个纳米的量级。
Resumo:
A deep binary silicon grating as high-extinction-ratio reflective polarizing beam splitter (PBS) at the wavelength of 1550 nm is presented. The design is based on the phenomenon of total internal reflection (TIR) by using the rigorous coupled wave analysis (RCWA). The extinction ratio of the rectangular PBS grating can reach 2.5×105 with the optimum grating period of 397 nm and groove depth of 1.092 μm. The effciencies of TM-polarized wave in the 0th order and TE-polarized wave in the −1st order can both reach unity at the Littrow angle. Holographic recording technology and inductively coupled plasma (ICP) etching could be used to fabricate the silicon PBS grating.
Resumo:
Tobacco BY-2 cells were exposed to microcystin-RR (MC-RR) at two concentrations, 60 mu g mL(-1) and 120 mu g mL(-1), to study the changes in morphology and ultrastructure of cells as a result of the exposure. Exposure to the lower concentration for 5 d led to typical apoptotic morphological changes including condensation of nuclear chromatin, creation of a characteristic 'half moon' structure, and cytoplasm shrinkage and decreased cell volume, as revealed through light microscopy. fluorescence microscopy, and transmission electron microscopy, respectively. Exposure to the higher concentration, on the other hand, led to morphological and ultrastructural changes typical of necrosis, such as rupture of the plasma membrane and the nuclear membrane and a marked swelling of cells. The presence of many vacuoles containing unusual deposits points to the involvement of vacuoles in detoxifying MC-RR. Results of the present study indicate that exposure of tobacco BY-2 cells to MC-RR at a lower concentration (60 mu g mL(-1)) results in apoptosis and that to a higher concentration (120 mu g mL(-1)), in necrosis. (C) 2009 Elsevier Ltd. All rights reserved.
Resumo:
Thymidylate synthase (TS), an essential enzyme in DNA synthesis and repair, plays a key role in the events of cell cycle regulation and tumor formation. Here, an investigation was presented about subcellular location and biological function of viral TS from lymphocystis disease virus from China (LCDV-C) in fish cells. Fluorescence microscopy revealed that LCDV-C TS was predominantly localized in the cytoplasm in fish cells. Cell cycle analysis demonstrated that LCDV-C TS promoted cell cycle progression into S and G2/M phase in the constitutive expressed cells. As a result, the cells have a faster growth rate compared with the control cells as revealed by cell growth curves. For foci assay, the TS-expressed cells gave rise to foci 4-5 weeks after incubation. Microscopic examination of the TS-induced foci revealed multilayered growth and crisscross morphology characteristic of transformed cells. Moreover, LCDV-C TS predisposed the transfected cells to acquire an anchorage-independent phenotype and could grow in 0.3% soft agar. So the data reveal LCDV-C TS is sufficient to induce a transformed phenotype in fish cells in vitro and exhibits its potential ability in cell transformation. To our knowledge, it is the first report on viral TS sequences associated with transforming activity. (C) 2007 Elsevier Inc. All rights reserved.