基于微球透镜的任选区高分辨光学显微成像新方法研究

提出和发展了基于毛细管-微球组合探针的任选区、 高分辨显微成像新方法. 建立了微球显微成像的物理模型, 利用成像理论,推导出微球成像的放大倍率; 采用3.0, 4.4, 5.6, 7.5, 10.0 μm等不同直径的SiO₂微球, 对未经刻录的DVD光盘进行了微球显微成像实验, 可以观察到DVD光盘的微纳米结构被明显放大且对比度显著提高, 与理论计算结果相符合; 采用毛细管微探针操纵微球的方法, 实现了基于微球透镜阵列的样品微纳米结构的高分辨显微成像; 在此基础上, 进一步将毛细管微探针与微球组合, 制备出毛细管-微球组合型探针, 首次实现了基于微球透镜的样品任意区域高分辨显微成像.     

Synthetic aperture fourier holographic optical microscopy

We report a new synthetic aperture optical microscopy in which high-resolution, wide-field amplitude and phase images are synthesized from a set of Fourier holograms. Each hologram records a region of the complex two-dimensional spatial frequency spectrum of an object, determined by the illumination field's spatial and spectral properties and the collection angle and solid angle. We demonstrate synthetic microscopic imaging in which spatial frequencies that are well outside the modulation transfer function of the collection optical system are recorded while maintaining the long working distance and wide field of view.     

Wide-field high-resolution structured illumination solid immersion fluorescence microscopy

The use of aplanatic solid immersion lenses (ASILs) made of high-refractive-index optical materials provides a route to wide-field high-resolution optical microscopy. Structured illumination microscopy (SIM) can double the spatial bandwidth of a microscope to also achieve high-resolution imaging. We investigate the combination of ASILs and SIM in fluorescence microscopy, which we call structured illumination solid immersion fluorescence microscopy (SISIM), to pursue a microscopic system with very large NA and high lateral resolution. We demonstrate that the combination can produce a wide-field high-resolution microscopic system with bandwidth corresponding to an NA of 3. Future developments of the SISIM system to make it achieve even higher resolution are proposed.     

Optical scatter imaging: subcellular morphometry in situ with Fourier filtering

We demonstrate a quantitative optical scatter imaging (OSI) technique, based on Fourier filtering, for detecting alterations in the size of particles with wavelength-scale dimensions. We generate our scatter image by taking the ratio of images collected at high and low numerical aperture in central dark-field microscopy. Such an image spatially encodes the ratio of wide to narrow angle scatter and hence provides a measure of local particle size. We validated OSI on sphere suspensions and live cells. In live cells, OSI revealed biochemically induced morphological changes that were not apparent in unprocessed differential interference contrast images. Unlike high-resolution imaging methods, OSI can provide size information for particles smaller than the camera's spatial resolution.     

High-resolution two-dimensional image upconversion of incoherent light

We consider a technique for high-resolution image upconversion of thermal light. Experimentally, we demonstrate cw upconversion with a resolution of more than 200 × 1000 pixels of thermally illuminated objects. This is the first demonstration (to our knowledge) of high-resolution cw image upconversion. The upconversion method promises an alternative route to high-quantum-efficiency all-optical imaging in the mid-IR wavelength region and beyond using standard CCD cameras. A particular advantage of CCD cameras compared to state-of-the-art thermal cameras is the possibility to tailor and tune the spectral response leading to functional spectral imaging.     

Hybrid division of aperture/division of a focal-plane polarimeter for real-time polarization imagery without an instantaneous field-of-view error

The direct, instantaneous measurement of the Stokes parameters of optical radiation is not possible using only a single detector. To infer the Stokes parameters in an image, the optical intensity must be modulated in time, space, or spectral characteristics and multiple measurements must be made on a pixel-by-pixel basis across the scene. Most existing imaging polarimetry strategies generally suffer from either spatial or temporal misregistration. Those strategies that do not suffer from misregistration have severe restrictions placed on the spectral properties of the illumination and/or optical system. A hybrid polarimeter is proposed that can be used for broadband imaging applications that, in principle, can be made without any temporal or spatial misalignment and that does not require detailed knowledge of the spectral properties of the radiation. The strategy uses arrays of microlenses and micropolarizers to create a high-resolution spatial array of nonimaging division of aperture polarimeters.     

Diffusion anisotropy in excised normal rat spinal cord measured by NMR microscopy

A conventional spin-echo NMR imaging pulse sequence was used to obtain high-resolution images of excised normal rat spinal cord at 7 and 14 T. It was observed that the large pulsed-field gradients necessary for high-resolution imaging caused a diffusion weighting that dominated the image contrast and that could be used to infer microscopic structural organization beyond that defined by the resolution of the image matrix (i.e., fiber orientation could be assigned based on diffusion anisotropy). Anisotropic diffusion coefficients were therefore measured using apparent diffusion tensor (ADT) imaging to assess more accurately fiber orientations in the spinal cord; structural anisotropy information is portrayed in the six unique images of the complete ADT. To reduce the dimensionality of the data, a trace image was generated using a separate color scale for each of the three diagonal element images of the ADT. This new image retains much of the invariance of the trace to the relative orientations of laboratory and sample axes (inherent to a greyscale trace image) but provides, by the use of color, contrast reflecting diffusion anisotropy. The colored trace image yields a pseudo-three-dimensional view of the rat spinal cord, from which it is possible to deduce fiber orientations.     

Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy

We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. The method involves acquiring two images, one with nonuniform illumination (in our case, speckle) and another with uniform illumination (in our case, randomized speckle). An evaluation of the local contrast in the speckle-illumination image provides an optically sectioned image with low resolution. This is complemented with high-resolution information obtained from the uniform-illumination image. A fusion of both images leads to a full resolution image that is optically sectioned across all spatial frequencies. This hybrid illumination method is fast, robust, and generalizable to a variety of illumination and imaging configurations.     

显微热成像系统自适应位置标定方法

利用光学微扫描技术,可在不改变探测器结构的情况下,提高显微热成像系统空间分辨率。但为了获得高质量过采样重构图像,微扫描位置需要标定。基于零点定标,提出一种各点自适应定标的微扫描位置标定方法。模拟实际系统在定标前后的欠采样图像,采用不同重构方法进行仿真对比验证;完成了实际采集的欠采样显微热图像序列的重构对比实验。实验结果表明该方法明显改善了显微热成像系统的过采样重构图像质量,提高了系统空间分辨率。此方法还可以应用在其他光电成像系统中。     

Toward two-dimensional nanometer resolution hard X-ray differential-interference-contrast imaging using modified photon sieves

In this Letter, we report a significant step forward in the design of single-optical-element optics for two-dimensional (2D) hard X-ray differential-interference-contrast (DIC) imaging based on modified photon sieves (MPSs). MPSs were obtained by a modified optic, i.e., combining two overlaid binary gratings and a photon sieve through two logical XOR operations. The superior performance of MPSs was demonstrated. Compared to Fresnel zone plates-based DIC diffractive optical elements (DOEs), which help to improve contrast only in one direction, MPSs can provide better resolution and 2D DIC imaging. Compared to normal photon sieves, MPSs are capable of imaging at a significantly higher image contrast. We anticipate that MPSs can provide a complementary and versatile high-resolution nondestructive imaging tool for ultra-large-scale integrated circuits at 45 nm node and below.     

Confocal enhanced optical coherence tomography for nondestructive evaluation of paints and coatings

We investigate confocal enhanced optical coherence tomography as a nondestructive evaluation tool for imaging with improved image contrast and depth resolution through a highly scattering paint layer. The depth responses provided by confocal microscopy, optical coherence tomography, and confocal enhanced optical coherence tomography for imaging through paint are investigated. We demonstrate that a new combination of confocal microscopy and optical coherence tomography provides significantly improved resolution and contrast for coherence gated imaging of substrate structures under the paint.     

激光焊接熔透特征信号同轴增效提取方法研究

熔透检测是实现高功率激光焊接质量在线控制的重要环节,但由于介观尺度下的低辐值熔透特征信号产生于激光匙孔底部被匙孔喷射物质和周围干扰信号完全掩盖,熔透状态难以被直接获取,常规检测多以间接测量为主。将光谱透视技术、红外显微成像技术、光电传感技术及空间定位提取技术相结合,提出一种激光焊接熔透特征信号同轴增效提取方法。以高功率激光在匙孔内壁激发的荧光辐射源作为直接检测信号,利用不同发光体的谱段特性在红外谱段有效分离并抑制激光焊接匙孔上方的等离子体、金属蒸汽焰、粒子团簇等强干扰信号,使红外荧光信号得到有效增强,实现光谱透视显像效果。同时采用自行研制的激光焊接同轴显微光路系统,利用红外显微成像原理提取到匙孔内壁受激辐射荧光的红外显微实像。并以此为基础对高功率激光焊接熔透状态与匙孔内部影像特征进行关联研究,发现与熔透状态直接相关的低辐射值特征现象及特征区域的存在。通过视觉辅助定位调节和熔透特征位置试验矫正等寻位方式,依次提高定位精度,直至将传感器光电感应芯片高精度定位至荧光辐射实像中的熔透特征区域。由此通过光谱透视-显微成像-介观寻位萃取的逐层光学分离方式,实现了对匙孔熔透特征数据的精准提取和最大化增强。试验结果表明,基于多种光谱及光学处理技术复合应用的大功率固体激光焊熔透特征同轴增效提取方法对激光熔透特征信号增强效果显著,可作为一种新型的高功率激光焊接熔透在线检测手段。     

基于液晶可调变滤色器的光谱图像获取及颜色重建方法

基于液晶可调变滤色器(LCTF)的光谱成像系统具有光谱分辨率高、结构紧凑、控制灵活等特点。搭建了一套可见光波段的高分辨率LCTF光谱成像系统,对系统的光谱图像获取及彩色图像重建方法进行了研究;建立了系统的光谱及颜色重建模型;基于标准色卡及标准测色系统,对该系统的颜色复现效果进行了实验验证;实验结果表明,该LCTF光谱成像系统的彩色图像颜色复现精度达可到较小色差水平。     

Spectral-domain optical coherence phase and multiphoton microscopy

We describe simultaneous quantitative phase contrast and multiphoton fluorescence imaging by combined spectral-domain optical coherence phase and multiphoton microscopy. The instrument employs two light sources for efficient optical coherence microscopic and multiphoton imaging and can generate structural and functional images of transparent specimens in the epidirection. Phase contrast imaging exhibits spatial and temporal phase stability in the subnanometer range. We also demonstrate the visualization of actin filaments in a fixed cell specimen, which is confirmed by simultaneous multiphoton fluorescence imaging.     

基于声光可调谐滤光器的显微光谱成像技术

为了解决传统声光可调谐滤光器(AOTF)成像模糊的缺点,设计了一种新的AOTF。该器件通过在传统的AOTF的出射孔后面放置一个自行设计的等边色散棱镜来实现对衍射光的色散展宽进行补偿,明显地提高了成像的对比度和空间分辨率。将此器件附加在传统光学显微镜上,获得了一种新型的光谱显微成像仪器。其光谱分辨率在575nm波长处为4.2nm、成像空间分辨率为2μm、图像采集速度为毫秒量级。为基于AOTF的光谱成像技术在生物医学等领域的更广泛的应用奠定了基础。     

光源尺寸和光谱带宽对波带板成像的影响

X射线菲涅耳波带板成像能实现亚微米空间分辨能力,有可能应用于激光等离子体或聚变靶的高分辨X射线成像诊断.之前的数值模拟研究表明,成像分辨能力受光源尺寸、入射光或成像光谱带宽的影响.本文报道在632.8 nm为中心波长的可见光波段,对波带板成像的数值模拟和原理性验证实验.数值模拟表明:随着扩展光源尺寸增加,视场中央分辨能力基本不变,而像对比度下降;随着成像的光谱带宽的增加,视场中央分辨能力与像对比度同时下降.实验证实了数值模拟的结论,且实验与数值模拟结果的定量比较也符合得较好.     

基于主成分分析-二阶导数光谱成像的红外显微图像分析

红外显微成像技术将红外光谱技术和显微技术相结合,不仅可以提供测试样品的光谱信息而且能够提供测试样品的空间分布信息。然而复杂样品的红外显微图像谱峰重叠严重,无法直接获得目标组分的分布信息。将因子分析与光谱分离技术相结合提出了主成分分析-二阶导数光谱成像方法。通过兔子动脉红外显微图像中胆固醇分布的成像实验,验证该方法的可行性和有效性。实验结果表明,该方法可以提高光谱分辨率,挖掘隐藏于重叠谱峰中的有用信息,是一种有效的红外显微图像分析方法。     

Three-dimensional visualization of objects in scattering medium using integral imaging and spectral analysis

This paper presents a three-dimensional visualization method of 3D objects in a scattering medium. The proposed method employs integral imaging and spectral analysis to improve the visual quality of 3D images. The images observed from 3D objects in the scattering medium such as turbid water suffer from image degradation due to scattering. The main reason is that the observed image signal is very weak compared with the scattering signal. Common image enhancement techniques including histogram equalization and contrast enhancement works improperly to overcome the problem. Thus, integral imaging that enables to integrate the weak signals from multiple images was discussed to improve image quality. In this paper, we apply spectral analysis to an integral imaging system such as the computational integral imaging reconstruction. Also, we introduce a signal model with a visibility parameter to analyze the scattering signal. The proposed method based on spectral analysis efficiently estimates the original signal and it is applied to elemental images. The visibility-enhanced elemental images are then used to reconstruct 3D images using a computational integral imaging reconstruction algorithm. To evaluate the proposed method, we perform the optical experiments for 3D objects in turbid water. The experimental results indicate that the proposed method outperforms the existing methods.     

光声信号的双谱分析方法研究

基于光声效应的光声光谱技术是光学与声学的有机结合, 可利用不同波长的入射光波, 产生不同的光声信号, 从而为组分识别、组织无损检测等提供高对比度图像, 是一种极具潜力的新型医学诊断技术.但光声光谱检测技术由于受检测方法的制约, 往往扫描时间较长, 而且信号的稳定性和图像识别的准确性也较差. 为了弥补单一光声光谱分析的局限性, 根据光声效应原理和振动理论, 提出了一种光声光谱与光声频谱相结合的双谱分析方法. 该方法通过光声频域信息的定量分析, 可以有效地提高不同组织的光声图像对比度, 从而提高光声成像的组分识别能力, 为光声频谱功能成像奠定理论基础. 实验结果显示, 光声光谱与光声频谱相结合的双谱分析方法可以较好地识别实验组织样品, 可实现高速、高分辨率的组分识别、组织探伤等, 具有广泛的应用前景和重要的临床诊断意义. 关键词： 光声成像 光声光谱分析 光声频谱分析     

深海光学照明与成像系统分析及进展

深海光学成像系统分为4个子系统:照明系统、相机系统、图像处理系统以及数据存储与传输系统,本文对深海光学成像系统化研究与发展趋势展开分析。文中有针对性地对深海光学成像最前端的两个子系统-照明系统与相机系统进行了较为详细地阐述。其中,深海照明系统进一步细分为3个更小的系统:光源系统、配光系统以及灯阵系统;对于深海相机系统则根据其应用领域及技术特点细分为水下普通成像、激光成像、偏振光成像、3D/全景成像、显微成像以及光谱成像6类。从近年来国内外深海光学成像的发展历史及现状来看,其未来的发展趋势可以归结为以下几点:更高的分辨率,更深的工作深度,更大的观测范围以及更多样的成像方式。