Spatial filter based 3D resolution improvement and polarization properties of multiphoton multiple-excitation-spot-optical microscopy

Three-dimensional (3D) resolution improvement in multi-photon multiple-excitation-spot-optical microscopy is proposed. Specially designed spatial filter is employed for improving the overall 3D resolution of the imaging system. An improvement up to a factor of 14.5 and sub-femto liter volume excitation is achieved. The system shows substantial sidelobe reduction (<4%) due to the non-linear intensity dependence of multiphoton process. Polarization effect on x-oriented and freely rotating dipoles shows dramatic change in the field distribution at the focal-plane. The resulting point-spread function has the ability to produce several strongly localized polarization dependent field patterns which may find applications in optical engineering and bioimaging.

Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy

Monitoring and visualizing specimens at a large penetration depth is a challenge. At depths of hundreds of microns, several physical effects (such as, scattering, PSF distortion and noise) deteriorate the image quality and prohibit a detailed study of key biological phenomena. In this study, we use a Bessel-like beam in-conjugation with an orthogonal detection system to achieve depth imaging. A Bessel-like penetrating diffractionless beam is generated by engineering the back-aperture of the excitation objective. The proposed excitation scheme allows continuous scanning by simply translating the detection PSF. This type of imaging system is beneficial for obtaining depth information from any desired specimen layer, including nano-particle tracking in thick tissue. As demonstrated by imaging the fluorescent polymer-tagged-CaCO3 particles and yeast cells in a tissue-like gel-matrix, the system offers a penetration depth that extends up to 650 mu m. This achievement will advance the field of fluorescence imaging and deep nano-particle tracking.

Spatial filter based light-sheet laser interference technique for three-dimensional nanolithography

We propose a laser interference technique for the fabrication of 3D nano-structures. This is possible with the introduction of specialized spatial filter in a 2 pi cylindrical lens system (consists of two opposing cylindrical lens sharing a common geometrical focus). The spatial filter at the back-aperture of a cylindrical lens gives rise to multiple light-sheet patterns. Two such interfering counter-propagating light-sheet pattern result in periodic 3D nano-pillar structure. This technique overcomes the existing slow point-by-point scanning, and has the ability to pattern selectively over a large volume. The proposed technique allows large-scale fabrication of periodic structures. Computational study shows a field-of-view (patterning volume) of approximately 12: 2mm(3) with the pillar-size of 80 nm and inter-pillar separation of 180 nm. Applications are in nano-waveguides, 3D nano-electronics, photonic crystals, and optical microscopy. (C) 2015 AIP Publishing LLC.

Spatial filtering nearly eliminates the side-lobes in single- and multi-photon 4pi-type-C super-resolution fluorescence microscopy

Super-resolution microscopy has tremendously progressed our understanding of cellular biophysics and biochemistry. Specifically, 4pi fluorescence microscopy technique stands out because of its axial super-resolution capability. All types of 4pi-microscopy techniques work well in conjugation with deconvolution techniques to get rid of artifacts due to side-lobes. In this regard, we propose a technique based on spatial filter in a 4pi-type-C confocal setup to get rid of these artifacts. Using a special spatial filter, we have reduced the depth-of-focus. Interference of two similar depth-of-focus beams in a 4 pi geometry result in substantial reduction of side-lobes. Studies show a reduction of side-lobes by 46% and 76% for single and two photon variant compared to 4pi - type - C confocal system. This is incredible considering the resolving capability of the existing 4pi - type - C confocal microscopy. Moreover, the main lobe is found to be 150 nm for the proposed spatial filtering technique as compared to 690 nm of the state-of-art confocal system. Reconstruction of experimentally obtained 2PE - 4pi data of green fluorescent protein (GFP)-tagged mitocondrial network shows near elimination of artifacts arising out of side-lobes. Proposed technique may find interesting application in fluorescence microscopy, nano-lithography, and cell biology. (C) 2013 AIP Publishing LLC.

Spatial coherence does not affect contrast discrimination for multiple Gabor stimuli

Gestalt grouping rules imply a process or mechanism for grouping together local features of an object into a perceptual whole. Several psychophysical experiments have been interpreted as evidence for constrained interactions between nearby spatial filter elements and this has led to the hypothesis that element linking might be mediated by these interactions. A common tacit assumption is that these interactions result in response modulation which disturbs a local contrast code. We addressed this possibility by performing contrast discrimination experiments using two-dimensional arrays of multiple Gabor patches arranged either (i) vertically, (ii) in circles (coherent conditions), or (iii) randomly (incoherent condition), as well as for a single Gabor patch. In each condition, contrast increments were applied to either the entire test stimulus (experiment 1) or a single patch whose position was cued (experiment 2). In experiment 3, the texture stimuli were reduced to a single contour by displaying only the central vertical strip. Performance was better for the multiple-patch conditions than for the single-patch condition, but whether the multiple-patch stimulus was coherent or not had no systematic effect on the results in any of the experiments. We conclude that constrained local interactions do not interfere with a local contrast code for our suprathreshold stimuli, suggesting that, in general, this is not the way in which element linking is achieved. The possibility that interactions are involved in enhancing the detectability of contour elements at threshold remains unchallenged by our experiments.

Scaling geospatial searches in large spatial databases

Modern geographical databases, which are at the core of geographic information systems (GIS), store a rich set of aspatial attributes in addition to geographic data. Typically, aspatial information comes in textual and numeric format. Retrieving information constrained on spatial and aspatial data from geodatabases provides GIS users the ability to perform more interesting spatial analyses, and for applications to support composite location-aware searches; for example, in a real estate database: “Find the nearest homes for sale to my current location that have backyard and whose prices are between $50,000 and $80,000”. Efficient processing of such queries require combined indexing strategies of multiple types of data. Existing spatial query engines commonly apply a two-filter approach (spatial filter followed by nonspatial filter, or viceversa), which can incur large performance overheads. On the other hand, more recently, the amount of geolocation data has grown rapidly in databases due in part to advances in geolocation technologies (e.g., GPS-enabled smartphones) that allow users to associate location data to objects or events. The latter poses potential data ingestion challenges of large data volumes for practical GIS databases. In this dissertation, we first show how indexing spatial data with R-trees (a typical data pre-processing task) can be scaled in MapReduce—a widely-adopted parallel programming model for data intensive problems. The evaluation of our algorithms in a Hadoop cluster showed close to linear scalability in building R-tree indexes. Subsequently, we develop efficient algorithms for processing spatial queries with aspatial conditions. Novel techniques for simultaneously indexing spatial with textual and numeric data are developed to that end. Experimental evaluations with real-world, large spatial datasets measured query response times within the sub-second range for most cases, and up to a few seconds for a small number of cases, which is reasonable for interactive applications. Overall, the previous results show that the MapReduce parallel model is suitable for indexing tasks in spatial databases, and the adequate combination of spatial and aspatial attribute indexes can attain acceptable response times for interactive spatial queries with constraints on aspatial data.

Fusion of morphological images for airborne target detection

Several track-before-detection approaches for image based aircraft detection have recently been examined in an important automated aircraft collision detection application. A particularly popular approach is a two stage processing paradigm which involves: a morphological spatial filter stage (which aims to emphasize the visual characteristics of targets) followed by a temporal or track filter stage (which aims to emphasize the temporal characteristics of targets). In this paper, we proposed new spot detection techniques for this two stage processing paradigm that fuse together raw and morphological images or fuse together various different morphological images (we call these approaches morphological reinforcement). On the basis of flight test data, the proposed morphological reinforcement operations are shown to offer superior signal to-noise characteristics when compared to standard spatial filter options (such as the close-minus-open and adaptive contour morphological operations). However, system operation characterised curves, which examine detection verses false alarm characteristics after both processing stages, illustrate that system performance is very data dependent.

Note: Dynamic point spread function for single and multiphoton fluorescence microscopy

We propose and demonstrate a dynamic point spread function (PSF) for single and multiphoton fluorescence microscopy. The goal is to generate a PSF whose shape and size can be maneuvered from highly localized to elongated one, thereby allowing shallow-to-depth excitation capability during active imaging. The PSF is obtained by utilizing specially designed spatial filter and dynamically altering the filter parameters. We predict potential applications in nanobioimaging and fluorescence microscopy.

Note: Dynamic point spread function for single and multiphoton fluorescence microscopy

We propose and demonstrate a dynamic point spread function (PSF) for single and multiphoton fluorescence microscopy. The goal is to generate a PSF whose shape and size can be maneuvered from highly localized to elongated one, thereby allowing shallow-to-depth excitation capability during active imaging. The PSF is obtained by utilizing specially designed spatial filter and dynamically altering the filter parameters. We predict potential applications in nanobioimaging and fluorescence microscopy.

Generation of extended light-sheets for single and multi-photon fluorescence microscopy

We theoretically propose and computationally demonstrate the generation of extended light-sheet for fluorescence microscopy. This is made possible by the introduction of a specially designed double-window spatial filter that allows the light to pass through the periphery and center of a cylindrical lens. When illuminated with a plane wave, the proposed filter results in an extended depth-of-focus along with side-lobes which are due to other interferences in the transverse focal plane. Computational studies show a maximum extension of light-sheet by 3.38 times for single photon excitation and 3.68 times for multiphoton excitation as compared to state-of-art single plane illumination microscopy system. This technique may facilitate the study of large biological specimens (such as Zebrafish embryo and tissue) with high spatial resolution and reduced photobleaching. (C) 2013 AIP Publishing LLC.

Three Dimensional Fluorescence Imaging Using Multiple Light-Sheet Microscopy

We developed a multiple light-sheet microscopy (MLSM) system capable of 3D fluorescence imaging. Employing spatial filter in the excitation arm of a SPIM system, we successfully generated multiple light-sheets. This improves upon the existing SPIM system and is capable of 3D volume imaging by simultaneously illuminating multiple planes in the sample. Theta detection geometry is employed for data acquisition from multiple specimen layers. This detection scheme inherits many advantages including, background reduction, cross-talk free fluorescence detection and high-resolution at long working distance. Using this technique, we generated 5 equi-intense light-sheets of thickness approximately 7: 5 mm with an inter-sheet separation of 15 mm. Moreover, the light-sheets generated by MLSM is found to be 2 times thinner than the state-of-art SPIM system. Imaging of fluorescently coated yeast cells of size 4 +/- 1 mm (encaged in Agarose gel-matrix) is achieved. Proposed imaging technique may accelerate the field of fluorescence microscopy, cell biology and biophotonics.

High-power extracavity pulse compression in solid materials

A novel technique for high-power extracavity pulse compression with a nonlinear solid material is demonstrated. Before spectral broadening by self-phase modulation in the solid material, a short filament generated in argon is used as a spatial filter, which works for a uniform spectrum broadening over the spatial profile. Compensated by chirped mirrors, a 15-fs pulse is generated from a 32-fs input laser pulse. A total transmission larger than 80% after the solid material is achieved.

The modulated grating hologram

A phase and amplitude, off-axis hologram has been synthesized from three computer-generated transmission masks, using a multiple-exposure holographic recording method. Each of the masks controls one fixed-phase component of the complex hologram transmittance. The basic grating is generated optically, relieving the computer of the burden of drawing details the size of each fringe. The maximum information capacity of the computer plotting device can then be applied to the generation of the grating modulation function. By this method large digital holograms (25 mm by 25 mm) have been synthesized in dichromated gelatin. The recording method is applicable to virtually any holographic medium.

The modulated grating hologram was designed primarily for the application of spatial filtering, in which the requirement is a hologram with large dynamic range and large free spectral range. Choice of a low-noise, high-efficiency medium such as dichromated gelatin will allow exceptionally large dynamic range. Independence of the optically-generated carrier grating from the computer-generated modulation functions allows arbitrarily large free spectral range.

The performance of a holographic spatial filter will be limited ultimately by noise originating from imperfections in the holographic medium. The characteristics of this noise are analyzed, and in the case of a high diffraction efficiency hologram are shown to differ significantly from previous analyses. The dominant noise source in holograms of high diffraction efficiency will be scattering of the first order or imaging wave by deformations in the hologram surface or other effects of low spatial frequency. Experimental measurements in various low-noise holographic media verify these predictions.

Laser beam automatic alignment in multipass amplifier

A laser beam automatic alignment system is applied in a multipass amplifier of the SG-III prototype laser. Considering the requirements of the SG-III prototype facility, by combining the general techniques of the laser beam automatic alignment system, according to the image relayed of the pinholes in the spatial filter, and utilizing the optical position and the spatial distribution of the four pinholes of the main spatial filter in the multipass amplifier of the SG-III prototype, a reasonable and optimized scheme for automatic aligning multipass beam paths is presented. It is demonstrated on the multipass amplifier experimental system. (C) 2004 Society of Photo-Optical Instrumentation Engineers.

高斯光束通过空间滤波器的传输特性

采用矩阵分解和用有限个复高斯函数之和逼近硬边光阑窗口函数相结合的方法，从柯林斯衍射积分公式出发，对高斯光束通过空间滤波器的传输特性进行了分析，得到了相应的解析式。通过对解析式的数值模拟，得出了直观的结果，并进行了误差分析。数值计算结果表明，最佳滤波小孔尺寸的选择和计算误差均与光束的截断参数有关。在此基础上，进一步研究了空间滤波器具有像传递功能时，高斯光束和平顶高斯光束（N=12）通过空间滤波器的传输特性，得出了截断参数与滤波后光强分布的关系，并证实了此方法也可用于计算其他类型光束通过空间滤波器的传输。