一种提高频谱编码成像技术信噪比的方法

频谱编码成像技术是一种采用衍射光栅把不同波长照明到样品的不同位置处的新型反射式显微成像技术。搭建了一个基于50 k Hz扫频光源的频谱编码显微系统,为解决无后置放大器情况下探测微弱样品光的问题,采用平衡探测的方法进行了成像测试。通过对USAF-1951分辨率板成像测得横向分辨率由13.93μm提高到5.52μm,采用平衡探测的方法使得洋葱样品图像信噪比(SNR)由15.07 d B提高到22.6 d B。研究结果表明,采用平衡探测的方法能够提高图像分辨率和信噪比。对离体猪胃小凹样品进行成像,验证了频谱编码成像技术在生物消化道内成像的可行性,为下一步该方法实现临床应用奠定了理论基础。     

一种提高频谱编码成像技术信噪比的方法EI北大核心CSCD

频谱编码成像技术是一种采用衍射光栅把不同波长照明到样品的不同位置处的新型反射式显微成像技术。搭建了一个基于50 k Hz扫频光源的频谱编码显微系统,为解决无后置放大器情况下探测微弱样品光的问题,采用平衡探测的方法进行了成像测试。通过对USAF-1951分辨率板成像测得横向分辨率由13.93μm提高到5.52μm,采用平衡探测的方法使得洋葱样品图像信噪比(SNR)由15.07 d B提高到22.6 d B。研究结果表明,采用平衡探测的方法能够提高图像分辨率和信噪比。对离体猪胃小凹样品进行成像,验证了频谱编码成像技术在生物消化道内成像的可行性,为下一步该方法实现临床应用奠定了理论基础。     

频谱编码内窥成像技术

频谱编码成像技术是一种利用光栅在横向位置编码的新型生物组织成像技术。该技术采用一个光栅和一个聚焦透镜在样品上产生光谱编码线,无需额外扫描便可得到一条线的图像,跟频域干涉技术相结合,再加上慢速的线扫描便可获得三维图像,在内窥成像上具有很大的实用价值。在分析频谱编码成像技术基本原理的基础上,推导了主要的性能参数。通过对分辨率测试靶进行成像,验证分析影响横向分辨率的因素。结果表明,编码线方向的分辨率与入射光斑直径成反比,与焦距成正比;垂直于编码线方向分辨率要弱于编码线方向的分辨率。最后对洋葱样品进行成像,验证了频谱编码成像技术的可行性。     

基于电动可调焦透镜的大范围快速光片显微成像

搭建了一种基于电动可调焦透镜(electrically tunable lens)的大范围快速光片荧光显微成像系统.通过引入电动可调焦透镜与一维振镜以实现成像物平面和光片位置的快速移动,再结合高速s CMOS完成快速光片荧光显微成像.另外实验中通过改善光路与提升动态成像质量,实现了大范围扫描并减少了伪像.最终对成像性能进行测试,本系统的纵向分辨率和横向分辨率分别达到约5.5μm和约0.7μm,单幅图像稳定成像的速度约为275 frames/s,成像深度可超过138μm,能满足对具有一定尺寸的生物样本进行实时清晰成像的需求.     

线照明并行谱域光学相干层析成像系统与缺陷检测应用研究

针对玻璃缺陷在线无损检测的迫切需求,本文报道了一种基于线照明并行谱域光学相干层析成像系统的大视场检测系统.该系统采用快速面阵CMOS相机,单次拍摄即可获取完整的横截面(B-scan)图像.基于线照明面阵探测器的并行谱域光学相干层析成像系统,可以同时获取沿线照明方向各位置处的深度分辨信息,避免了横向扫描机构的应用.研制系统的轴向分辨率为17.9μm,并行方向上的横向分辨率55.7μm,扫描方向上的横向分辨率为24.8μm,轴向扫描速率为128 000 A-scan/s,横向视场为32 mm,空气中成像深度大于6 mm,成像灵敏度达到62 dB以上.利用研制的线照明并行谱域光学相干层析成像系统,开展了不同类型玻璃表面及其内部缺陷的检测应用研究.     

基于菲涅耳透镜色散的双波长荧光显微深度成像

提出一种非轴向扫描的细胞显微深度成像技术,在显微系统中加入菲涅耳透镜,利用菲涅耳透镜的色散将不同激发光波长聚焦到不同的轴向位置,以实现对两个或多个焦平面同时成像.基于405nm和532nm两种激发光波长,在传统的荧光显微镜的激发路径中加入对应的两个成像探测器来探测两个不同焦平面所对应像面的成像信息,搭建得到一个能够实现探测深度约为12μm的基于菲涅耳透镜的荧光显微深度成像系统,并与基于显微物镜色差无菲涅耳透镜的荧光显微深度成像系统的成像深度和轴向分辨率进行实验对比.实验结果表明:加入菲涅耳透镜能够实现系统对不同焦面的同时成像;对于同一荧光波段,保证系统横向分辨率的同时,扩大了成像景深.该系统可以实现荧光生物细胞内部不同深度处的多波段同时探测.     

小型光谱编码显微成像系统的研制

光谱编码显微镜使用一个衍射光栅和光谱分析装置来获得显微图像。样品上不同的位置被不同的波长照明,通过对反射光光谱进行解码来得到空间信息。研制了一个基于扫频光源和平衡探测器的小型光谱编码显微镜(CSEM)。在没有放大器的情况下,一个固定增益的平衡探测器被用来探测较弱的样品光。通过对一个1951USAF分辨率测试靶成像来测量系统的横向分辨率。对离体猪小肠组织和在体手指皮肤成像来验证生物组织成像性能。结果表明,CSEM具有对生物组织深度分辨成像的能力。     

基于液体变焦透镜和振镜的三维光片显微成像系统

搭建了一种基于液体变焦透镜和振镜的三维光片显微成像系统,设计了振镜、液体变焦透镜、相机的同步控制采集成像系统,通过调谐振镜和液体变焦透镜,使得光片激发样品和成像同步,获得样品不同切面的图像堆栈并实现样品的三维重建。当采用数值孔径为0.3、放大倍率为10的成像物镜时,该系统的轴向扫描范围为507μm,横向视场达到1970μm×1300μm,横向分辨率为1.32μm,轴向分辨率可达12.75μm。在轴向扫描过程中,系统的放大倍率保持恒定,可以用于对一定尺寸生物样品的成像实验和相关研究,并通过对斑马鱼胚胎进行成像验证所提系统对厚生物样品成像的可行性。     

内窥式扫频光源光学相干层析系统的探头设计

利用自聚焦透镜和单模光纤搭建了基于光纤旋转连接器的内窥式扫频光源光学相干层析成像系统.利用Zemax模拟了内窥探头的光学性能,分析了光纤到自聚焦透镜的距离和自聚焦透镜节距对探头性能参量的影响.在综合考虑元器件成本以及成像要求后,选择无需定制,节距为0.24Pitch的自聚焦透镜,确定光纤与自聚焦透镜之间的距离为0.7mm.经实验测得该系统的横向分辨率约为19.5μm,纵向分辨率约为9μm,工作距离约为7mm,成像深度为6.2mm(空气中),与理论值接近.为了验证该系统对生物组织的成像性能,利用该系统对猪食道进行离体成像,重建后的层析图中可以明显地观察到猪肠道的表皮层和固有层.测量系统的各项成像性能参量以及对生物组织的成像性能表明该探头在内窥成像中是可行的.     

硅基半导体芯片激光红外显微成像系统研究

为了解决可见光显微成像技术无法实现硅基芯片内部结构观测的问题,根据1 064nm的红外激光对硅材料具有一定穿透深度的特性,设计了一种硅基半导体芯片激光红外显微成像系统.该系统采用数值孔径为0.42的长工作距显微物镜,通过音圈电机振动多模石英光纤消除散斑噪音,由系统观察CD-RW盘片道间距,表明系统分辨率可达到1.6μm,接近理论值,实现了对芯片厚度为70μm的静态随机存储器内部结构显微成像的观测.     

Reflection-mode photoacoustic microscopy using a hollow focused ultrasound transducer for in vivo imaging of blood vessels

A reflection-mode photoacoustic microscope using a hollow focused ultrasound transducer is developed for highresolution in vivo imaging.A confocal structure of the laser and the ultrasound is used to improve the system resolution.The axial and lateral resolutions of the system are measured to be ～ 32 μm and ～ 58 μm,respectively.Ex vivo and in vivo modes are tested to validate the imaging capability of the photoacoustic microscope.The adjacent vein and artery can be seen clearly from the reconstructed photoacoustic images.The results demonstrate that the reflectionmode photoacoustic microscope can be used for high-resolution imaging of micro-blood vessels,which would be of great benefit for monitoring the neovascularization in tumor angiogenesis.     

超高分辨光学相干层析成像技术与材料检测应用

本文报道了一种超高分辨率谱域光学相干层析成像(SD-OCT)系统. 该系统基于超连续谱激光光源并截取部分光谱作为宽带光源, 其中心波长为665 nm, 光谱半高全宽(FWHM) 230 nm. 系统轴向分辨率0.9 μm, 轴向扫描速率28600行/秒, 横向分辨率3.9 μm, 横向视场1 mm, 最大成像深度0.6 mm(空气中). 利用研制的超高分辨率SD-OCT系统, 对不同型号的工业砂纸精细结构进行了成像, 并与普通SD-OCT的成像结果进行对比, 充分展示了研制系统在材料无损检测中优势.     

双层塑料靶丸的X射线相衬成像

 为对微聚焦X射线相衬成像技术应用于双层以及多层塑料靶丸成像和特征分析进行可行性研究,基于类同轴X射线成像技术,综合考虑成像放大倍数、分辨率和衬度等因素,选择合适的实验参数,成功获得了较为清晰的双层塑料靶丸X射线相衬成像照片;采用数字图像处理技术的图像分割手段,如拉普拉斯高斯边缘检测法等对所成像中靶丸边界特征进行分析处理,获得了双层靶丸内层厚度为（10.5±0.6） μm,外层厚度为（9.2±0.7） μm,靶丸外径为（273.3±1.0） μm等参数。     

用于活体人眼视网膜观察的自适应光学成像系统

利用自适应光学技术,研制了两套活体人眼视网膜高分辨力成像系统,在实时校正人眼波前误差的基础上,实现活体人眼视网膜细胞尺度的高分辨力成像。这两套系统分别采用19和37单元小型压电变形反射镜作为波前校正元件,哈特曼-夏克(Hartmann-Shack)波前传感器测量波前误差,用眼底反射的半导体激光作为波前探测的信标。在用计算机控制自适应光学系统实现人眼波前误差校正后,触发闪光灯照明视网膜,用CCD相机记录视网膜的高分辨力图像。校正后的残余波前误差的均方根值已分别小于1／6和1／10波长,相当于视网膜上成像分辨力分别为3．4μm和2．6μm,接近衍射极限。试验表明37单元系统的成像质量更好。     

Multifocal multiphoton endoscope

We report a miniaturized resonant/non-resonant multi-fiber raster scanner that is paired with a gradient-index lens assembly to achieve a compact and flexible multifocal multiphoton endoscope capable of longitudinal parallel image acquisition. Multiphoton images are obtained simultaneously at three axial depths, separated by ≥4.8 μm, by incorporating three axially offset double clad optical fibers into the miniaturized scanner. The fabricated endoscope has an outer diameter of 3 mm, a rigid length of 4 cm, and acquires images at 4 frames/s per focal plane, with lateral and axial resolutions for two-photon imaging of 0.8 and 10 μm, respectively.     

Small angle X-ray scattering by spherical particles of Polystyrene and Polyvinyltoluene

The high intensity, high resolution multiple reflection diffractometer has been applied to make accurate measurements of the small angle X-ray scattering of eight different Dow Chemical latexes with stated diameters 2.051, 1.305, 0.814, 0.557, 0.365, 0.264, 0.126, and 0.088 μm. Using thin dry samples, the measured intensities of diffraction extrema with orders higher than the third were found to agree well with the simple Rayleigh-Gans theory which neglects multiple scattering as well as interparticle interference. Orders below the fourth showed a distinct effect of interparticle interference. This effect could be demonstrated to vanish when a liquid solution sample was used instead of a dry one. Scattering curves of thick dry samples had strongly decreased contrast between maxima and minima, an effect which is known to be due to multiple scattering. The following particle diameters were determined from the measured scattering curves: 2.102±0.013 μm, 1.313±0.008 μm, 0.823±0.004 μm, 0.552±0.002 μm, 0.352±0.002 μm, 0.254±0.002 μm, 0.119±0.001 μm and 0.078±0.001 μm. The discrepancies between these and the stated sizes are larger for the smaller particles. It is believed that the X-ray measurements are more accurate than the stated sizes which are based on measurements in the electron microscope. Therefore, as secondary standards, the diameter obtained from the X-ray measurements should be used.     

Quadrature fringes wide-field optical coherence tomography and its applications to biological tissues

We propose and demonstrate quadrature fringes wide-field optical coherence tomography (QF WF OCT) to expand an optical Hilbert transformation to two-dimensions. This OCT simultaneously measures two quadrature interference images using a single InGaAs CCD camera to obtain en face OCT images. The axial and lateral resolutions are measured at 29 μm in air and 70 μm limited by a pixel size of camera using a superluminescent diode with a wavelength of 1.3 μm as the light source; the system sensitivity is determined to be −90 dB. The area of the en face OCT images is 4.0 mm × 4.0 mm (160 × 160  pixels). The OCT images are measured axially with steps of 10 μm. The en face OCT images of a in vivo human fingertip and a in situ rat brain are three-dimensionally measured up to the depth of about 3 mm with some degradations of a lateral resolution.     

Label-free 3D imaging of microstructure, blood, and lymphatic vessels within tissue beds in vivo

This Letter reports the use of an ultrahigh resolution optical microangiography (OMAG) system for simultaneous 3D imaging of microstructure and lymphatic and blood vessels without the use of an exogenous contrast agent. An automatic algorithm is developed to segment the lymphatic vessels from the microstructural images based on the fact that the lymph fluid is optically transparent. An OMAG system is developed that utilizes a broadband supercontinuum light source, providing an axial resolution of 2.3 μm and lateral resolution of 5.8 μm, capable of resolving the capillary vasculature and lymphatic vessels innervating microcirculatory tissue beds. Experimental demonstration is performed by showing detailed 3D lymphatic and blood vessel maps, coupled with morphology, within mouse ears in vivo.     

Super-Resolution Two-Photon Fluorescence Tomography Through the Phase-Shifted Optical Beatings of Bessel Beams for High-Resolution Deeper Tissue 3D Imaging

A novel z-scanning-free epi-detected super-resolution two-photon fluorescence tomography (TPFT) technique enabling super-resolution deeper tissue 3D imaging is reported. To accomplish this, a unique method is conceived by generating the phase-shifted optical beatings of Bessel beams (PS-OB³) with a spatial light modulator (SLM) to break the diffraction limit for enhancing both the lateral and axial resolutions as well as improving the penetration depth in TPFT for super-resolution deeper tissue imaging. By electronically varying the optical beating frequency and the phase shifts of the beating patterns through SLM, the depth-resolved TPF signals about the volumetric tissue are encoded in the spatial frequency domain and hence, a series of depth-resolved TPF images can be retrieved by implementing inverse fast Fourier transform without a need of mechanical depth-scanning. PS-OB³ TPFT provides ≈1.3- and 2-fold improvements in lateral and axial resolutions in comparison with conventional point-scan TPF imaging. It is also illustrated that the epi-detected PS-OB³ TPFT imaging with inherent scattering-resilient properties of the Bessel beams employed gives over 2-fold improvement in imaging depth in porcine brain tissue compared to conventional point-scan Gaussian beam TPF imaging. The z-scanning-free optical sectioning ability of PS-OB³ method developed in TPFT is universal, which can be readily extended to practically any other nonlinear optical imaging modalities for super-resolution deeper 3D imaging in biological and biomedical tissues.