Diffraction microtomography with sample rotation: influence of a missing apple core in the recorded frequency space

Diffraction microtomography in coherent light is foreseen as a promising technique to image transparent living samples in three dimensions without staining. Contrary to conventional microscopy with incoherent light, which gives morphological information only, diffraction microtomography makes it possible to obtain the complex optical refractive index of the observed sample by mapping a three-dimensional support in the spatial frequency domain. The technique can be implemented in two configurations, namely, by varying the sample illumination with a fixed sample or by rotating the sample using a fixed illumination. In the literature, only the former method was described in detail. In this report, we precisely derive the three-dimensional frequency support that can be mapped by the sample rotation configuration. We found that, within the first-order Born approximation, the volume of the frequency domain that can be mapped exhibits a missing part, the shape of which resembles that of an apple core. The projection of the diffracted waves in the frequency space onto the set of sphere caps covered by the sample rotation does not allow for a complete mapping of the frequency along the axis of rotation due to the finite radius of the sphere caps. We present simulations of the effects of this missing information on the reconstruction of ideal objects.      

360 degree realistic 3D image display and image processing from real objects

A 360-degree realistic 3D image display system based on direct light scanning method, so-called Holo-Table has been introduced in this paper. High-density directional continuous 3D motion images can be displayed easily with only one spatial light modulator. Using the holographic screen as the beam deflector, 360-degree full horizontal viewing angle was achieved. As an accompany part of the system, CMOS camera based image acquisition platform was built to feed the display engine, which can take a full 360-degree continuous imaging of the sample at the center. Customized image processing techniques such as scaling, rotation, format transformation were also developed and embedded into the system control software platform. In the end several samples were imaged to demonstrate the capability of our system.     

Two-wave mixing and energy transfer in BaTiO3 application to laser beamsteering

Energy transfer between the two recording beams in a BaTiO₃ photorefractive crystal is analyzed as a function of the following parameters: incident beams ratio, spatial frequency, pump beam intensity. Exponential gain coefficients Γ ≈ 20 cm^-1 are reached for optimized holographic recording conditions. Application of the energy transfer to a new method of laser beam deflection is proposed.     

Holographic contouring using double-source technique and Fourier transform analysis

This paper describes a double-exposure holographic contouring technique by the movement of the object beam through fibre optics. Single-mode optic fibres are used to transfer both object and reference beams. Between two exposures the object illumination beam is displaced. The contouring fringes can be visualised on the object surfaces. The fringe patterns are automatically processed by the fast Fourier transform (FFT) method, which produces a 3D perspective plot. Examples are given for different objects.     

样品转动方式对中子全息成像结果的影响

在中子全息成像实验中, 为避免透射中子干扰, 入射中子束方向与样品-探测器方向不能在一条直线上, 同时采用探测器移动结合样品转动或仅样品转动的方式, 避免探测系统的大范围移动. 因此在样品的转动过程中, 入射中子束和探测器相对于样品的位置同时发生改变, 内源全息项和内探测器全息项作为变量被记录在同一幅全息图中, 并在重建过程中相互干扰. 本文对基于中子三轴谱仪和四圆谱仪的三种不同转动方式进行了中子全息记录和重建模拟研究, 并讨论其修正方法. 结果表明, 各转动方式都可以通过适当的方法消除或减轻相关影响, 而其中基于三轴谱仪的纯样品转动方式可以使用两个探测器记录的方式, 避开入射中子束方向与样品-探测器方向不能在一条直线上的限制, 得到完整的全息图, 同时通过数据处理能基本消除相对转动造成的干扰, 达到理想的重建结果, 在条件允许的情况下应予优先采用.     

空间光调制器曲面拼接实现全息三维显示视角拓展

本文对多空间光调制器不同拼接方式拓展全息三维再现像视角的方法进行了分析,基于多片空间光调制器拼接拓展视角的思想,利用平面反射镜、分光镜和两片透射式空间光调制器设计了曲面拼接系统,进行了全息三维再现像的视角拓展实验研究。用该系统对四棱锥物体的层析菲涅尔衍射全息图进行再现,结果表明,总视角由基于单片空间光调制器的1.7°增大到3.2°,即拓展到约1.9倍,分光镜能够消除两片空间光调制器间的间隙,实现无缝拼接。该方法同样适用于更多空间光调制器的曲面拼接中,可以有效地拓展全息再现像的视场角大小。     

Spatial Fourier-decomposition optical fluorescen tomography-theoretical investigation

A new three-dimensional (3D) optical fluorescent tomographic imaging scheme is proposed with structured illumination and spatial Fourierdomain decomposition methods for the first time. In this spatial Fourier-decomposition optical fluorescence tomography (SF-OFT), the intensity of focused excitation light from an objective lens is modulated to be a cosine function along the optical axis of the system. For a given position in a two-dimensional (2D) raster scanning process, the spatial frequency of the cosine function along the optical axis sweeps in a proper range while a series of fluorescence intensity are detected accordingly. By making an inverse discrete cosine transformation of these recorded intensity profiles, the distribution of fluorescent markers along the optical axis of a focused laser beam is obtained. A 3D optical fluorescent tomography can be achieved with this proposed SF-OFT technique with a simple 2D raster scanning process.     

Conjugate Image Plane Correlator with Holographic Disk Memory

A conjugate image plane correlator with holographic disk memory is proposed. Optical correlation between conjugate images reconstructed from a holographic disk and an input image on liquid-crystal television is executed with the rotation of the disk. Regardless of Fourier hologram recording with the pseudorandom diffuser, it is found possible to take out the diffuser from the original hologram recording scheme using an image reconstruction process and to get correlation signals between input and reconstructed conjugate images in the output plane of a two-lens imaging system. Generation of conjugate replicas with high contrast causes exact matching with an input image which results in high recognition performance for autocorrelation signals. The transfer function of an optical system can be controlled by adjustment of either hologram size or hologram area illuminated with a laser beam. Hence, the output intensity distribution can be adjusted by selecting a proper pupil function and the size of an output pupil defined by the input pupil size and the optical system magnification factor. The real-time character recognition by optical parallel high-speed processing for two dimensional images with position normalization is demonstrated.     

稀疏孔径双圆周阵列的成像性能研究

针对传统稀疏孔径均匀圆周阵列的调制传递甬数在截止频率区域内易出现零值而影响成像质量的问题,提出一种由两个均匀圆周阵列嵌套构成的无冗余的双圆周稀疏孔径结构.给出该阵列的光瞳函数以及调制传递函数的解析表达式.以5-3结构的双圆周阵列为例,以实际空间截止频率为像质评价准则,从子孔径直径、内环圆周半径以及内环旋转角度等几个方面详细分析了双圆周阵列的成像特性,给出不同情况下的实际截止频率值,并对双圆周阵列稀疏孔径系统进行模拟成像和图像复原.将该结构与均匀圆周阵列.Comwell优化圆周阵列进行比较.结果表明,相同条件下,该阵列具有更大的实际空间截止频率,并且成像质量随着填充因子的减小而逐渐下降.     

Three-dimensional optical transfer of rotating beams

Systems in which the point spread function (PSF) is a rotating beam have increasing use in three-dimensional (3D) microscopy and depth estimation. We analyze in several ways the 3D optical transfer function (OTF) of Gauss Laguerre modes and rotating beams. This is based on analysis of 3D OTFs of general aperture functions. Consequently, we suggest a criterion for depth resolution based on an effective cutoff of the axial frequency response. This criterion can be used to optimize PSFs explicitly and directly, to maximize axial resolution.     

基于实际空间截止频率的稀疏孔径双圆周阵列的优化

提出了一种新的稀疏孔径光瞳结构-两个圆周阵内外嵌套而成的双圆周结构,并研究了该结构的调制传递函数.该结构以实际空间截止频率作为约束条件,保证调制传递函数在最小截止频率以内无零频,以阵列的u-v覆盖点间距最大化及最小冗余度来设计目标函数,并采用这种优化方法对3-3、5-3以及6-3双圆周阵列进行了优化排列.将优化结果与相同子孔径数目的均匀圆周阵以及Golay结构进行仿真实验和比较.结果表明,优化后的双圆周阵列,具有最大的实际空间截止频率和实际等效口径.     

基于涡旋光照明的暗场数字全息显微方法研究

提出了一种基于涡旋光照明的暗场数字全息显微方法. 从理论上阐述了涡旋光环形照明原理和暗场数字全息显微原理, 分析了涡旋光的准无衍射特性对成像的影响; 搭建了相应的数字全息显微成像系统, 采用690 nm的聚苯乙烯小球作为实验样品; 最后通过对小球明暗场下数字全息显微再现像的分析对比, 证明该方法可以有效地提高数字全息系统的分辨率, 同时增强了再现像的对比度. 关键词： 全息 暗场数字全息显微 涡旋光 分辨率     

Holography with local reference beam obtained by spatial frequency separation

The holographic method disclosed in this paper allows holographic recording with a very high reference-to-object wave energy ratio without attenuating the rays diffracted by the object. In those holographic methods permitting local generation of the reference beam, which have been proposed so far, the reference beams are obtained by either wavefront or amplitude division of the wave of coherent radiation impinging onto the optical apparatus. It is readily seen that most of the incident energy is used to process the reference beam reducing drastically the energy of the object bearing wave. In the novel method proposed here the reference wave is obtained by separation of the low spatial frequency components from the high-order spatial frequency components which bear the object information. As a result of the removal of the low frequency components, a dark-field image is recorded and later reconstructed by the holographic plate. Even though the configuration is of the off-axis type, proper design may reduce the coherence requirements of the source of radiation to those for in-line holography. The paper discusses the basic properties of the method and experimental results are presented. This work forms part of a dissertation to be submitted by M. L. A. Gassend in partial fulfillment of the requirements for the Ph.D. degree. It has been supported by the National Research Council of Canada under Grants Nos. A 7240 and T 0409, and by the Department of External Affairs (Award No. 759193 administered by the Canada Council).     

基于实际空间截止频率的稀疏孔径双圆周阵列的优化

提出了一种新的稀疏孔径光瞳结构—两个圆周阵内外嵌套而成的双圆周结构,并研究了该结构的调制传递函数.该结构以实际空间截止频率作为约束条件,保证调制传递函数在最小截止频率以内无零频,以阵列的u－v覆盖点间距最大化及最小冗余度来设计目标函数,并采用这种优化方法对3-3、5-3以及6-3双圆周阵列进行了优化排列.将优化结果与相同子孔径数目的均匀圆周阵以及Golay结构进行仿真实验和比较.结果表明,优化后的双圆周阵列,具有最大的实际空间截止频率和实际等效口径.     

荧光波长对共焦显微镜成像特性的影响

导出了共焦显微镜中不同荧光波长情况下的荧光功率传输函数、三维脉冲响应函数（3D－PSF）和三维光学传递函数（3D－OTF）。结果表明，不同的荧光波长对共焦显微镜的空间截止频率、分辨率、光学传递函数存在明显的影响。当激发波长与荧光波长的比值下降到一定程度时，可以看到明显的失锥现象。     

Improved and isotropic resolution in tomographic diffractive microscopy combining sample and illumination rotation

Tomographic Diffractive Microscopy is a technique, which permits to image transparent living specimens in three dimensions without staining. It is commonly implemented in two configurations, by either rotating the sample illumination keeping the specimen fixed, or by rotating the sample using a fixed illumination. Under the first-order Born approximation, the volume of the frequency domain that can be mapped with the rotating illumination method has the shape of a “doughnut”, which exhibits a so-called “missing cone” of non-captured frequencies, responsible for the strong resolution anisotropy characteristic of transmission microscopes. When rotating the sample, the resolution is almost isotropic, but the set of captured frequencies still exhibits a missing part, the shape of which resembles that of an apple core. Furthermore, its maximal extension is reduced compared to tomography with rotating illumination. We propose various configurations for tomographic diffractive microscopy, which combine both approaches, and aim at obtaining a high and isotropic resolution. We illustrate with simulations the expected imaging performances of these configurations.     

基于单一分光棱镜干涉仪的双通路定量相位显微术

仅仅使用一个单独的分光棱镜(BS),实现了一种用于生物细胞三维成像的双通路定量相位显微术.不同于传统的使用方法,将BS倾斜放置,使中央半反射层与入射光光轴之间存在一个非常小的角度.这样基于BS的分光特性,经过BS后的透射光束和反射光束将会叠加在一起并形成干涉.调节样品位置,利用相机拍摄同时获得了存在π相移的双通路干涉图.这种离轴干涉模式,只需要记录单幅干涉图就可以获得真实的相位信息,方法结构简单,易于操作,适用于微小透明样品的三维形貌测量.     

Rotation of single live mammalian cells using dynamic holographic optical tweezers

We report on a method for rotating single mammalian cells about an axis perpendicular to the optical system axis through the imaging plane using dynamic holographic optical tweezers (HOTs). Two optical traps are created on the opposite edges of a mammalian cell and are continuously transitioned through the imaging plane along the circumference of the cell in opposite directions, thus providing the torque to rotate the cell in a controlled fashion. The method enables a complete 360° rotation of live single mammalian cells with spherical or near-to spherical shape in 3D space, and represents a useful tool suitable for the single cell analysis field, including tomographic imaging.     

Driving and analysis of micro-objects by digital holographic microscope in microfluidics

We propose an optical configuration in which floating particles in a microfluidic chamber can be characterized by an interference microscopy configuration to obtain quantitative phase-contrast maps. The configuration is simply made by two laser beams from the same laser source. One beam provides the optical forces for driving the particle along appropriate paths, but at same time works as the object illumination beam in the holographic microscope. The second beam plays the role of the reference beam, allowing recording of an interference fringe pattern (i.e., the digital hologram) in an out-of-focus image plane. The system and method are illustrated and experimental results are offered for polymeric particles as well as for in vitro cells with the aim to demonstrate the approach.     

Investigation of in-plane stresses on bolted flange joints using digital speckle pattern interferometry

Digital Speckle Pattern Interferometry (DSPI) is a promising field for a wide range of applications such as measurement of displacements/deformations, contouring, nondestructive testing, etc. It has holographic sensitivity and circumvents delays in photographic processing/filtering. Speckle correlation interferometry can be made sensitive to only one of the components of in-plane displacement by illuminating the object with two beams symmetric about the viewing axis and imaging along this axis using a viction. This configuration allows measurement of in-plane displacements independent of out-of-plane displacements.

In this paper, the in-plane deformation/stress distribution has been studied for a typical bolted flange joint of a pressure vessel using two beam illumination DSPI. The information obtained can be very useful in predicting the leakage behaviour of such joints employed in pressure vessels. Experimental investigations along with theoretical predictions are presented.