Focal plane tuning in wide-field-of-view microscope with Talbot pattern illumination

We have developed a focal plane tuning technique for use in focus-grid-based wide-field-of-view microscopy (WFM). In WFM, the incidence of a collimated beam on a mask with a two-dimensional grid of aperture produced the Talbot images of the aperture grid. The Talbot pattern functioned as a focus grid and was used to illuminate the sample. By scanning the sample across the focus grid and collecting the transmission, we can generate a microscopy image of the sample. By tuning the wavelength of the laser, we can tune the focal plane of the WFM and acquire images of different depth into the sample. Images of a green algae microscope slide were acquired at different focal planes for demonstration.     

用于大景深单分子定位显微的多功能全息相位片的设计及数值模拟

发展具有大轴向定位范围的单分子定位技术对于实现厚样品的超分辨成像具有重要的价值.基于波前编码技术,将变形多值纯相位光栅与双螺旋点扩散函数相位片相结合,提出一种可以通过空间光调制器实现的具有高衍射效率的新型全息相位片的设计方法.这种全息相位片可以将样品内多个层面的分子信息以双螺旋的形式成像在同一个探测面的不同位置,在无需扫描的情况下提高双螺旋点扩散函数工程的轴向定位范围和分辨率,解决活细胞内单分子定位和示踪技术中的大景深探测难题.数值模拟表明,设计的5×5全息相位片可以将样品内25个层面上的分子信息以双螺旋的形式成像在同一探测面上的不同位置,相邻两个层面的间隔为0.5μm,实现了轴向12μm的探测范围,证明了设计的可行性.     

Improved depth resolution in video-rate line-scanning multiphoton microscopy using temporal focusing

By introducing spatiotemporal pulse shaping techniques to multiphoton microscopy it is possible to obtain video-rate images with depth resolution similar to point-by-point scanning multiphoton microscopy while mechanically scanning in only one dimension. This is achieved by temporal focusing of the illumination pulse: The pulsed excitation field is compressed as it propagates through the sample, reaching its shortest duration (and highest peak intensity) at the focal plane before stretching again beyond it. This method is applied to produce, in a simple and scalable setup, video-rate two-photon excitation fluorescence images of Drosophila egg chambers with nearly 100,000 effective pixels and 1.5 microm depth resolution.     

基于相位展开及拼接算法的一种高精度大量程宽光谱干涉显微术

宽光谱干涉显微术广泛应用于高精密检测领域, 它测量样品形貌通常采用垂直扫描干涉术对亚微米至毫米级特征进行测量,以及相移干涉术对纳米级特征进行测量。其中,相移干涉术精度可达纳米级,但量程有限,高度变化对应的相位需限制在区间内。采用包裹相位展开算法可以扩展相移干涉术的量程,也仅适用于平滑表面,当高度起伏超出焦深或者光源相干长度的限定范围时,干涉条纹模糊或对比度丧失,所解算的结果将产生较大误差甚至错误。提出一种基于相位展开及拼接算法的高精度、大量程宽光谱干涉显微测量方法,以干涉条纹调制度量化条纹质量,条纹对比度高、成像清晰的区域对应调制度较高,定义当前焦面条纹调制度高于阈值的区域为理想区域,定义焦面条纹调制度低于阈值的区域为问题区域。以相位展开算法获得理想区域中的样品相位分布,问题区域的包裹相位不进行展开。使用微位移结构纵向移动物镜焦平面,选择合理的步长,使相邻焦面位置理想区域展开后的真实相位保持部分区域重合,根据重合区域的相位值均差可以实现不同焦面位置的高精度相位拼接,最终获得扩展量程的高精度真实相位结果,进而可以恢复样品完整的表面形貌分布。该算法通过对理想区域的筛选,避免了相位在问题区域展开带来的误差,可以得到精确的测量结果。通过模拟计算和实验验证,证明了该方法不仅保持了宽光谱干涉显微术中相移干涉术的纳米级高精度,还可将其量程从数百纳米拓展到数微米。而且,该方法精度不依赖于位移部件,理论上量程可以拓展到显微物镜的全工作距离。     

波前编码显微物镜景深扩展的研究

为了解决传统显微物镜景深与分辨率的矛盾,采用波前编码的方法,设计研究了波前编码的10倍显微物镜,结合传统光学设计软件,采用基于MTF一致性的相位板参数的优化方法,实现了在像面附近一定范围内系统的点扩散函数一致性;此外,还设计了扩展三次相位编码板的10倍显微物镜,比较了采用两种不同相位板系统的焦深扩展的效果,结果显示三次相位板的焦深扩展效果较好,可以将传统10倍显微物镜的焦深扩展15倍。成像模拟仿真结果显示滤波后的编码像在±15倍焦深范围内成像清晰,从而扩大了系统的景深。     

Simultaneous imaging of multiple focal planes using a two-photon scanning microscope

Despite all the advances in nonlinear microscopy, all existing instruments are constrained to obtain images of one focal plane at a time. In this Letter we demonstrate a two-photon absorption fluorescence scanning microscope capable of imaging two focal planes simultaneously. This is accomplished by temporally demultiplexing the signal coming from two focal volumes at different sample depths. The scheme can be extended to three or more focal planes.     

聚焦高斯涡旋光束焦区电场和磁场的偏振奇点

以高斯涡旋光束为例,研究了聚焦场中电场和磁场的偏振奇点变化规律.结果表明,高斯涡旋光束经透镜聚焦后,在焦区存在二维和三维电场和磁场的偏振奇点,其位置一般不重合.适当改变与焦面的距离以及光阑截断参数等出现圆偏振奇点的移动、产生和湮没.不同二维和三维电场和磁场圆偏振奇点湮没所对应的各参数临界值不同.在二维电场中,几何焦面上会有V点的产生.     

Resolution improvement in optical projection tomography by the focal scanning method

Optical projection tomography (OPT) requires the depth of field (DOF) of the lens to cover at least half of the sample. There is a trade-off between obtaining high resolution with a high-NA lens and obtaining large DOF with a low-NA lens. The DOF of a high-NA objective lens can be extended by scanning its focal plane through the sample. We call this extended DOF image a "pseudoprojection." Images reconstructed from these pseudoprojections have isometric resolution, which can be the same as the lateral resolution of the high-NA objective. The focal scanning method produces an over 10× improvement in OPT resolution.     

Continuous-wave terahertz digital holography by use of a pyroelectric array camera

Terahertz (THz) digital holography is realized based on a 2.52?THz far-IR gas laser and a commercial 124 × 124 pyroelectric array camera. Off-axis THz holograms are obtained by recording interference patterns between light passing through the sample and the reference wave. A numerical reconstruction process is performed to obtain the field distribution at the object surface. Different targets were imaged to test the system's imaging capability. Compared with THz focal plane images, the image quality of the reconstructed images are improved a lot. The results show that the system's imaging resolution can reach at least 0.4 mm. The system also has the potential for real-time imaging application. This study confirms that digital holography is a promising technique for real-time, high-resolution THz imaging, which has extensive application prospects.     

一种三维物体相息图的迭代计算方法

在传统迭代傅里叶变换算法的基础上,提出了一种计算三维物体相息图的新方法.基于层析法将三维物体的多个分层物面作为衍射再现图像,在一个输入面(相息图)和多个输出面(再现像)之间进行迭代.通过在傅里叶迭代运算中引入距离相位因子,表示物体不同物面的深度,体现了物体的三维特征.实验结果证明了本文算法良好的收敛特性和再现性能.最后,分析了物面数量和间距对全息再现质量的影响,利用液晶空间光调制器采用时分复用的方法还原了三维物体的多个物面.     

表面高分辨弹性反冲探测分析

在中国原子能科学研究院 HI- 1 3串列加速器上建立了用 Q3D磁谱仪动量分析和ΔE- E粒子分辨对材料表面进行高分辨的弹性反冲探测分析技术 .用 1 0 0 Me V12 7I对 C/Li F多层样品的深度分布分析表明 ,表面分辨达到 1 .2 nm.所建立的ΔE(气体 ) - E(半导体 )望远镜探测器可同时分析从轻至中重的所有元素 .实测了新光电材料 Ga N,La2 Sr Cu O4 超导膜和新超硬材料 C3N4 (Si)等样品. High resolution depth profiling technique with elastic recoil detection analysis has been developed at the HI 13 tandem accelerator of CIAE. A depth resolution of 1.2 nm was achieved at the surface of the samples with the Q3D magnetic spectrometer and the focal plane detector. From light to medium heavy elements were simultaneous analyzed with a small Δ E E telescope. The method was applied to depth profile analysis of C/LiF multilayers, La 2SrCuO 4 superconductor and GaN foil samples.     

Depth-control method for integral imaging

We present a method for controlling the depth of three-dimensional (3D) images reconstructed by integral photography. Incoherent light is reflected from 3D objects, propagates through a lens array, and is captured as the first elemental images by a capturing device. The second elemental images of the 3D images are generated by numerical processing from the first elemental images in accordance with the desired depth. The optical reconstruction of 3D images at the desired depth by the second elemental images is confirmed experimentally.     

Surface normal imaging with a hand-held NMR device

Recently the capabilities of single sided nuclear magnetic resonance (NMR) devices have been extended towards three-dimensional imaging. This paper details the use of a magnetic field sweep coil to obtain spatial resolution in the plane normal to the surface of a hand-held NMR device-the NMR-Mobile Universal Surface Explorer (MOUSE). One-dimensional depth profiles can be recorded by varying the current in the sweep coils. Preliminary results from multi-layer rubber and glass sample phantoms demonstrate a sample penetration depth of 7 mm. Two-dimensional images were acquired via the inclusion of phase encoding coils. Non-destructive cross-sectional images of small rubber phantoms were successfully recorded.     

Dipole‐like backscatter stimulated Raman scattering for in vivo imaging

Stimulated Raman scattering (SRS) scanning microscopy has the potential to enable label‐free in vivo imaging for research and clinical medicine. Volume SRS from focus occurs in the forward scattered direction. Therefore, multiple scattering events are required to direct the light out of the tissue, reducing imaging depth and resolution. Here, a method called Stokes interference SRS (SISRS) is introduced that operates by the addition to the standard pump and stimulated emission probe beams a third beam called the donut beam. The donut is close in wavelength to the probe beam and, after passage through a π phase plate, forms an annular beam in the focal plane with bright nodes above and below focus. The donut beats with the probe beam, and when they destructively interfere with each other, the microscope's 3‐D stimulated emission focal spot is reduced to subwavelength dimensions. A subwavelength focal volume emits a dipole pattern of SRS with forward and backscatter lobes, enabling high‐resolution single‐backscatter imaging from deep within tissues. The reduction of the focal volume also increases the resolution of the scanning image creating imaging beyond the diffraction limit. SISRS imaging may provide in vivo label‐free Raman images comparable with that achieved in stained in vitro tissues in all planes of section. Copyright © 2014 John Wiley & Sons, Ltd.     

一种分焦平面偏振成像系统光源标定方法北大核心CSCD

水下偏振成像技术是目前水下成像研究的热点,由于自然光在水中衰减大,水下成像系统多采用主动照明方式。针对分焦平面偏振成像系统中偏振照明光源与偏振探测像元偏振方向不匹配引起采集图像偏振信息存在的偏差,进而影响目标图像增强质量的问题,提出了一种分焦平面偏振成像系统光源标定方法。阐述了偏振光源的标定原理,然后给出偏振光源标定的实施步骤,最后采用偏振去雾算法和图像质量评价方法对标定前后的水下目标图像进行了图像增强和图像质量评价。评价结果表明,标定后的增强图像质量优于未标定的增强图像质量,平均梯度最大提升了2.48倍。该标定方法简单有效,实用性强,适用于分焦平面偏振成像系统偏振光源标定。     

Common-path phase-shifting interferometer with binary grating

An experimental setup for phase extraction of 2D phase distributions is presented. The system uses a common-path interferometer consisting of two windows in the input plane and a translating grating as spatial filter. In the output, interference of the fields associated with replicated images of the input windows is achieved by a proper choice of the windows spacing with respect to the grating period, the focal length of the transforming lens and the wavelength of the coherent illumination employed. Because in this type of grating interferometer a grating is placed as a spatial filter, the phase changes which are needed for phase-shifting interferometry can be easily performed with translations of the grating driven by a linear actuator. Some experimental results are shown.     

Agitation-free multiphoton microscopy of oblique planes

The scanning two-photon fluorescence microscope produces optically sectioned images from the focal plane. It is sometimes desirable to acquire images from other planes of the specimen that are inclined with respect to the focal plane. In this Letter, we discuss the issues concerned with acquiring such images together with the effects of the inclination angle on image resolution and sectioning strength. To obtain images from oblique planes at high speed, a two-photon system was built wherein a novel optical system is used to provide aberration-free scanning.     

Arbitrary structuring of two-dimensional photonic crystals by use of phase-only Fourier gratings

The computer-generated holography technique is applied to the structuring of two-dimensional (2D) photonic crystals with inherently embedded arbitrary defects. The technique uses phase-only Fourier gratings as a generator of spot arrays in the focal plane, such that a single exposure produces a 2D array of focused spots with desired defects or modifications in the lattice structure. We demonstrate several types of large-area 2D lattice structures with square, hexagonal, or hybrid lattices embedded with point and (or) line defects. Scanning the Fourier plane in the depth direction throughout multiphoton polymerization media allows 3D lattices with stacked defect layers to be formed.     

Towards full-resolution inline electron holography

With the availability of fast computers, inline electron holography, a technique for reconstructing both amplitude and phase of the electron wave function as scattered by the sample from a set of differently aberrated transmission electron microscopy images, is becoming increasingly quantitative. While focal series reconstruction from transmission electron microscopy images has already been practiced for at least 3 decades, existing approaches can only recover a relatively small band of spatial frequencies. Here I present a reconstruction scheme which is capable of reconstructing the electron wave function for a very large range of spatial frequencies, demonstrating its performance using simulated as well as experimental data.     

畴腐蚀掺镁铌酸锂可调阵列光分束器的研究

研究了不同畴腐蚀深度的掺镁铌酸锂二维六角可调阵列光分束器的分数Talbot效应.对不同Talbot分数β和不同畴腐蚀深度的阵列光分束器Talbot衍射像进行了数值模拟理论研究.模拟结果表明,Talbot分数β可以改变Talbot衍射像的周期及结构分布,而畴腐蚀深度可有效调制衍射像的光强分布.在理论研究的基础上,设计并制备了具有不同畴腐蚀深度的掺镁铌酸锂二维六角阵列光分束器,对其在不同Talbot分数β条件下的分数Talbot效应进行了通光实验研究,实现了畴腐蚀阵列光分束器对近场Talbot衍射光强分布的调制,实验结果与理论研究结果一致.