Total-internal-reflection fluorescence microscopy with W-shaped axicon mirrors

A scheme based on a W-shaped axicon mirror device for total-internal-reflection fluorescence microscopy (TIRFM) is presented. This approach combines the advantages of higher efficiency compared with traditional TIRFM, adjustable illumination area, and simple switching between wide-field and TIRF imaging modes. TIRF images obtained with this approach are free of shadow artifacts and of interference fringes. Example micrographs of fluorescently labeled polystyrene beads, of Convallaria majalis tissue, and of Propidium-iodide-labeled Chinese hamster ovary cells are shown, and the capabilities of the scheme are discussed. (C) 2010 Optical Society of America

Biosensor with Total Internal reflection Imaging Ellipsometry

In order to monitor multiple protein reaction processes simultaneously, a biosensor based on imaging ellipsometry operated in the total internal reflection mode is proposed. It could be realised as an automatic analysis for protein interaction processes with real-time label-free method. Its principle and methodology as well as a demonstration for its applications are presented.

Constitutive plasma membrane targeting and microdomain localization of Dok5 studied by single-molecule microscopy

In the present study, single-molecule fluorescence microscopy was used to examine the characteristics of plasma membrane targeting and microdomain localization of enhanced yellow fluorescent protein (eYFP)-tagged wild-type Dok5 and its variants in living Chinese hamster ovary (CHO) cells. We found that Dok5 can target constitutively to the plasma membrane, and the PH domain is essential for this process. Furthermore, single-molecule trajectories analysis revealed that Dok5 can constitutively partition into microdomain on the plasma membrane. Finally, the potential mechanism of microdomain localization of Dok5 was discussed. This study provided insights into the characteristics of plasma membrane targeting and microdomain localization of Dok5 in living CHO cells. (C) 2008 Elsevier B.V. All rights reserved.

Nonpolarizing beam splitter designed by frustrated total internal reflection inside a glass cube

A method for the design of an all-dielectric nonpolarizing prism beam splitter utilizing the principle of frustrated total internal reflection is reported. The nonpolarizing condition for a prism beam splitter is discussed, and some single layer design examples are elaborated. The concept can be applied to a wide range of wavelengths and arbitrary transmittance values, and with the help of a computer design program examples of 400-700 nm, T-p = T-p = 0.5 +/- 0.01, with incident angles of 45 degrees and 62 degrees are given. In addition, the sensitivity and application of the design are also discussed. (c) 2006 Optical Society of America.

Design of high-efficiency diffraction gratings based on total internal reflection for pulse compressor

We present designs of high-efficiency compression grating based on total internal reflection (TIR) for picosecond pulse laser at 1053 nm. The setup is devised by directly etching gratings into the bottom side of a prism so that light can successfully enter (or exit) the compression grating. Dependence of the -1 order diffraction efficiencies on the constructive parameters is analyzed for TE- and TM-polarized incident light at Littrow angle by using Fourier modal method in order to obtain optimal grating structure. The electric field enhancement within the high-efficiency TIR gratings is regarded as another criterion to optimize the structure of the TIR gratings. With the criterion of high diffraction efficiency, low electric field enhancement and sufficient manufacturing latitude, TIR compression gratings with optimized constructive parameters are obtained for TE- and TM-polarized incident light, respectively. The grating for TE-polarized light exhibits diffraction efficiencies higher than 0.95 within 23 nm bandwidth and relatively low square of electric field enhancement ratio of 5.7. Regardless of the internal electric field enhancement, the grating for TM-polarized light provides diffraction efficiencies higher than 0.95 within 42 nm bandwidth. With compact structure, such TIR compression gratings made solely of fused silica should be of great interest for application to chirped pulse amplification (CPA) systems. (c) 2007 Elsevier B.V. All rights reserved.

Integrated folding 4x4 optical matrix switch with total internal reflection mirrors on SOI by anisotropic chemical etching

A folding rearrangeable nonblocking 4 x 4 optical matrix switch was designed and fabricated on silicon-on-insulator wafer. To compress chip size, switch elements (SEs) were interconnected by total internal reflection (TIR) mirrors instead of conventional S-bends. For obtaining smooth interfaces, potassium hydroxide anisotropic chemical etching of silicon was utilized to make the matrix switch for the first time. The device has a compact size of 20 x 1.6 mm(2) and a fast response of 7.5 mu s. The power consumption of each 2 x 2 SE and the average excess loss per mirror were 145 mW and -1.1 dB, respectively. Low path dependence of +/- 0.7 dB in total excess loss was obtained because of the symmetry of propagation paths in this novel matrix switch.

Novel folding large-scale optical switch matrix with total internal reflection mirrors on silicon-on-insulator by anisotropy chemical etching

A compact optical switch matrix was designed, in which light circuits were folded by total internal reflective (TIR) mirrors. Two key elements, 2 x 2 switch and TIR mirror, have been fabricated on silicon-on-insulator wafer by anisotropy chemical etching. The 2 x 2 switch showed very low power consumption of 140 mW and a very high speed of 8 +/- 1 mus. An improved design for the TIR mirror was developed, and the fabricated mirror with smooth and vertical reflective facet showed low excess loss of 0.7 +/- 0.3 dB at 1.55 mum.

SOI optical switch matrix integrated with spot size converter (SSC) and total internal reflection (TIR) mirrors

SOI (Silicon on Insulator) based photonic devices has attracted more and more attention in the recent years. Integration of SOI optical switch matrix with isolating grooves, total internal reflection (TIR) mirrors and spot size converter (SSC) was studied. A folding re-arrangeable non-blocking 4x4 optical switch matrix and a blocking 16x16 matrix with TIR mirrors and SSC were fabricated on SOI wafer. The performaces, including extinction ratio and the crosstalk, are better than before. The insertion loss and the polarization dependent loss (PDL) at 1.55 mu m increase slightly with longer device length, more bend and intersecting waveguides. The insertion losses decrease 2 similar to 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.

Giant Goos-Hanchen displacement enhanced by dielectric film in frustrated total internal reflection configuration

It is predicted that the Goos-Hanchen displacement in the usual frustrated total internal reflection configuration can be resonantly enhanced greatly by coating a dielectric thin film onto the surface of the first prism when the angle of incidence is larger than the critical angle for total reflection at the prism-vacuum interface and is smaller than but close to the critical angle for total reflection at the prism-film interface. Theoretical analysis shows that the displacement of transmitted beam is about half the displacement of reflected beam in the thick limit of the vacuum gap between the two prisms. This is to be compared with the relation in the usual symmetric double-prism configuration that the displacement of transmitted beam is equal to the displacement of reflected beam. Numerical simulations for a Gaussian incident beam of waist width of 100 wavelengths reveal that when the dielectric thin film is of the order of wavelength in thickness, both the reflected and transmitted beams maintain well the shape of the incident beam in the thick limit of the vacuum gap. So largely enhanced displacements would lead to applications in optical devices and integrated optics. (c) 2007 American Institute of Physics.

Large positive and negative lateral displacements from total internal reflection configuration with a weakly absorbing dielectric film

It is theoretically shown that the simultaneously large positive and negative lateral displacements will appear when the resonant condition is satisfied for a TE-polarized light beam reflected from the total internal reflection configuration with a weakly absorbing dielectric film. Appearance of the enhanced negative lateral displacement is relative to the incidence angle, absorption of the thin Elm and its thickness. If we select an appropriate weakly absorbing dielectric film and its thickness, the simultaneously enhanced positive and negative lateral displacements will appear at different resonant angles. These phenomena may lead to convenient measurements and interesting applications in optical devices.

The adsorption of protein molecules at a crystal/solution interface observed by an improved TIRFM

We have improved the ordinary total internal reflection fluorescence microscopy (TIRFM). Two improvements have been achieved, one is the interface between opaque material and solution can be observed, another is the interface far away (usually several ten micro meters) the objective lens can be observed. By this improved TIRFM, the adsorption of protein molecules at a crystal/solution interface had been successfully observed. We have obtained the results of relationship between the amount of adsorbed protein molecules on bunched steps and the height of bunched steps of a protein crystal.

Direct visualization of the dynamics of membrane-anchor proteins in living cells

Dynamic properties of proteins have crucial roles in understanding protein function and molecular mechanism within cells. In this paper, we combined total internal reflection fluorescence microscopy with oblique illumination fluorescence microscopy to observe directly the movement and localization of membrane-anchored green fluorescence proteins in living cells. Total internal reflect illumination allowed the observation of proteins in the cell membrane of living cells since the penetrate depth could be adjusted to about 80 nm, and oblique illumination allowed the observation of proteins both in the cytoplasm and apical membrane, which made this combination a promising tool to investigate the dynamics of proteins through the whole cell. Not only individual protein molecule tracks have been analyzed quantitatively but also cumulative probability distribution function analysis of ensemble trajectories has been done to reveal the mobility of proteins. Finally, single particle tracking has acted as a compensation for single molecule tracking. All the results exhibited green fluorescence protein dynamics within cytoplasm, on the membrane and from cytoplasm to plasma membrane.

离焦系统下的单分子三维取向研究

提出了一种新思路来确定单分子的三维取向,在全内反射荧光显微成像的基础上,在显微镜的油镜上加一个水层,水和油的折射率不同造成光程差,产生离焦,利用全内反射产生隐失波激发了单分子的纵向分量,并在探测器前增加了一个分光镜,把光路分成偏振方向相互垂直的两束光,使得探测器上的光场分布包含单分子三维取向的信息。推导了隐失场激发的偶极矩光场在探测器上的光强分布表达式,通过分析偏振方向相互垂直的两束光在探测器上的不同的强度分布,来判断单分子偶极矩的三维取向。

Imaging and tracking single fluorescent nanobeads in solution

In single-particle tracking (SPT), fluorescence video microscopy is used to record the motion images of single particle or single molecule. Here, by using a total-internal-reflection microscope equipped with an argon ion laser and a charge-coupled device (CCD) camera with high-speed and high-sensitivity, video images of single nanobeads in solutions were obtained. From the trajectories, the diffusion coefficient of individual nanobead was determined by the mean square displacements as a function of time. The sizes of nanobeads were calculated by Stokes-Einstein equation, and the results were compared with the actual values.