Optical diagnostics of mercury jet for an intense proton target

An optical diagnostic system is designed and constructed for imaging a free mercury jet interacting with a high intensity proton beam in a pulsed high-field solenoid magnet. The optical imaging system employs a backilluminated, laser shadow photography technique. Object illumination and image capture are transmitted through radiation-hard multimode optical fibers and flexible coherent imaging fibers. A retroreflected illumination design allows the entire passive imaging system to fit inside the bore of the solenoid magnet. A sequence of synchronized short laser light pulses are used to freeze the transient events, and the images are recorded by several high speed charge coupled devices. Quantitative and qualitative data analysis using image processing based on probability approach is described. The characteristics of free mercury jet as a high power target for beam-jet interaction at various levels of the magnetic induction field is reported in this paper.     

激光主动成像图像的多帧后处理算法研究

在确定激光器的发散角度、脉冲峰值功率、接收光学系统的参数以及确定激光器和接收光学系统的几何配置后,激光主动成像系统要想得到更远距离更高分辨率的微弱目标的图像,就只受微弱目标的探测成像及处理技术的制约,因为在较长的距离和有限的激光能量下,不可能立刻照明整个目标场景,接收成像端CCD上每个像素接收到的光照度也不可能达到相当的水平。为了在特定的脉冲激光能量下增加成像系统的成像距离,同时减轻大气扰动对成像分辨率的影响,研究了一种独特的图像处理算法,用激光多脉冲来获取整个目标场景的图像,采用辐射量来确定每次曝光时间内最大景物照明区域,这种独特的多帧后处理算法,可以在大气扰动和连续散粒噪声影响下,得到比传统连续照明方式更高分辨率的图像。     

Optical microscopy with improved resolution using two-beam interference of low-coherence light

In recent years, high-resolution microscopy using structured illumination has been practically applied for fluorescent bio-imaging. However, there is a large amount of speckle noise in reflected- and scattered-light images, because structured illumination is typically generated by laser-beam interference. Hence, this high-resolution imaging technique cannot be effectively used in industrial applications. In this study, we attempted to generate structured illumination using two-beam interference of low-coherence light for high-resolution and low-speckle imaging. First, we constructed an optical system consisting of a Michelson interferometer configured in such a manner that it achieved zero optical path-length difference and allowed the interference fringes to be manipulated. Then, we confirmed that the generated structured illumination width corresponded to the coherence length of the light source. As a final result of the resolution improvement experiment, the narrow sample pitch of 0.4 μm was successfully resolved beyond the diffraction limit of 0.74 μm with relatively less speckle noise.     

近红外距离选通成像系统作用距离研究

近红外脉冲激光照明的距离选通成像技术能够有效降低后向散射的影响,提高夜间以及恶劣环境下光电成像系统的作用距离。针对近红外激光距离选通成像系统的设计需要,以系统输出信噪比大于该目标的形状和识别概率所对应的人眼阈值信噪比为原理。在微光成像系统作用距离公式的基础上。添加主动照明系统的影响因素,建立了近红外激光距离选通成像系统的作用距离模型。该模型得到的结果和实际系统实验结果相比具有很好的一致性,表明该模型合理,可靠。在近红外激光距离选通成像系统的设计过程中,可以用此模型对系统进行作用距离预测。     

在线图像可视铁谱成像系统的像面照度均匀性

为了定量评价在线图像可视铁谱(OLVF)成像系统的像面照度均匀性,建立了一种像面照度模型。以像方参数及放大倍率表征物方视场,将物方视场区域离散化,采用朗伯余弦理论建立入瞳模式的像面照度模型,实现了像面照度的计算与均匀性评价。利用Matlab进行了物面照度仿真分析,确定了OLVF成像系统环形阵列光源的发光二极管(LED)数量,基于像面照度分析确定了最佳成像焦距和放大倍率。计算了油腔通油情况下成像系统中的光能量损耗以及磨粒沉积面的照度分布,建立了油液吸光系数与CCD像面轴上像点照度的关系。结果显示:LED发光强度已知时,仿真计算的像面不均匀度约为5.60%,实际测试的像面不均匀度为8%~9%,满足不均匀度≤10%的要求。开展了磨粒铁谱图像采集实验,结果表明:图像中磨粒清晰可辨,便于图像分割与视觉特征提取。提出的模型可定量描述OLVF成像系统的像面照度,可作为优化系统结构,提高系统成像性能的依据。     

Optical characterization of probes for photon scanning tunnelling microscopy

The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the sub-wavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however, difficult to evaluate in a direct manner and has most often been inferred from the apparent quality of recorded optical images. Complicated near-field optical imaging characteristics, together with the possibility of topographically induced artefacts, however, has increased demands for a more reliable probe characterization technique. Here we present experimental results obtained for optical characterization of two different probes by imaging of a well-specified near-field intensity distribution at various spatial frequencies. In particular, we observe that a sharply pointed dielectric probe can be highly suitable for imaging when using p -polarized light for the illumination. We conclude that the proposed scheme can be used directly for probe characterization and, subsequently, for determination of an optical transfer function, which would allow one to deduce from an experimentally obtained image of a weakly scattering sample the field distribution existing near the sample surface in the absence of the probe.     

Optical characterization of probes for photon scanning tunnelling microscopy

The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the subwavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however, difficult to evaluate in a direct manner and has most often been inferred from the apparent quality of recorded optical images. Complicated near-field optical imaging characteristics, together with the possibility of topographically induced artefacts, however, has increased demands for a more reliable probe characterization technique. Here we present experimental results obtained for optical characterization of two different probes by imaging of a well-specified near-field intensity distribution at various spatial frequencies. In particular, we observe that a sharply pointed dielectric probe can be highly suitable for imaging when using p-polarized light for the illumination. We conclude that the proposed scheme can be used directly for probe characterization and, subsequently, for determination of an optical transfer function. which would allow one to deduce from an experimentally obtained image of a weakly scattering sample the field distribution existing near the sample surface in the absence of the probe.     

Second‐harmonic microscopy of ex vivo porcine corneas

The ex vivo cornea of porcine eyes has been studied with second‐harmonic microscopy with a laboratory‐built system to examine the structure of collagen fibrils at different length scales, as well as the image dependence on polarization and wavelength of the illumination source. We found that collagen fibrils can effectively be visualized with second‐harmonic microscopy, in agreement with previous findings, at different wavelengths of the illumination. The same laser source used for imaging may also be used to induce changes to the corneal tissues that are observable both in the linear and second‐harmonic imaging channels. Such studies are essential first steps towards a future high‐resolution optical characterization technique for simultaneous corneal surgery and wound healing of the human eye.     

视网膜细胞显微镜的照明系统

为满足视网膜细胞成像照明光束的高亮度、窄谱宽要求,提出了一种用于视网膜细胞成像的照明系统,将半导体激光器发出的632.8nm激光耦合到芯径为105μm的多模光纤中,用聚光镜将多模光纤的出光端汇聚到一个旋转毛玻璃上,再用一个投射物镜将毛玻璃上的光源像投射到眼睛里照亮视网膜。试验发现该照明方法很好地消除了激光散斑,满足视网膜显微成像对高亮度光源的要求,光源有较窄的谱宽,成像系统色差很小,成像质量优于采用传统氙灯做照明光源的图像。该照明系统能较好地满足视网膜显微成像。     

复合式多点测量速度干涉仪光学系统设计

为满足爆炸冲击作用下物质界面的速度测量需求,设计了一种复合式多点测量的速度干涉仪。采用物方和像方双远心光路,将光纤阵列出射的照明激光定点投射到待测物面上,实现了针状滴注式照明,充分利用了照明激光能量,且保证了待测物面在运动过程中具有恒定的照度。成像系统像面采用末端为大芯径的锥形光纤接收信号光,既保证了物面运动过程中信号光与光纤的有效耦合,又保证了信号的单模输出,以便进入单模光纤马赫-曾德干涉仪进行差频干涉。采用具有微小楔角、沿直径方向镀矩形带状45°反射膜的反射镜,将照明光路与成像光路同轴,并校正了成像系统的大量像散。该干涉测量系统在物面运动10mm的行程中,物面滴注式照明照度保持恒定,像面光斑大小没有超出大芯径的光纤芯径。此光学系统能够满足爆炸冲击界面的大行程速度测量需求。     

基于成像式照度探测的菲涅尔光学助降系统检测方法研究

为了解决菲涅尔光学助降系统(FLOLS)的检测问题,文中根据FLOLS检测应满足的技术指标,提出了一种基于成像式照度探测法的菲涅尔灯发光强度测量方案,并根据分析设计研制了成像式照度探测装置。通过试验,结果表明该装置可远距离对大口径、大发光角度的菲涅尔灯进行发光强度的探测,满足菲涅尔光学助降系统检测装置测量的要求。     

Adaptive optics in spinning disk microscopy: improved contrast and brightness by a simple and fast method

Multiconfocal microscopy gives a good compromise between fast imaging and reasonable resolution. However, the low intensity of live fluorescent emitters is a major limitation to this technique. Aberrations induced by the optical setup, especially the mismatch of the refractive index and the biological sample itself, distort the point spread function and further reduce the amount of detected photons. Altogether, this leads to impaired image quality, preventing accurate analysis of molecular processes in biological samples and imaging deep in the sample. The amount of detected fluorescence can be improved with adaptive optics. Here, we used a compact adaptive optics module (adaptive optics box for sectioning optical microscopy), which was specifically designed for spinning disk confocal microscopy. The module overcomes undesired anomalies by correcting for most of the aberrations in confocal imaging. Existing aberration detection methods require prior illumination, which bleaches the sample. To avoid multiple exposures of the sample, we established an experimental model describing the depth dependence of major aberrations. This model allows us to correct for those aberrations when performing a z‐stack, gradually increasing the amplitude of the correction with depth. It does not require illumination of the sample for aberration detection, thus minimizing photobleaching and phototoxicity. With this model, we improved both signal‐to‐background ratio and image contrast. Here, we present comparative studies on a variety of biological samples.     

Time-domain whole-field fluorescence lifetime imaging with optical sectioning

A whole-field time-domain fluorescence lifetime imaging (FLIM) microscope with the capability to perform optical sectioning is described. The excitation source is a mode-locked Ti:Sapphire laser that is regeneratively amplified and frequency doubled to 415 nm. Time-gated fluorescence intensity images at increasing delays after excitation are acquired using a gated microchannel plate image intensifier combined with an intensified CCD camera. By fitting a single or multiple exponential decay to each pixel in the field of view of the time-gated images, 2-D FLIM maps are obtained for each component of the fluorescence lifetime. This FLIM instrument was demonstrated to exhibit a temporal discrimination of better than 10 ps. It has been applied to chemically specific imaging, quantitative imaging of concentration ratios of mixed fluorophores and quantitative imaging of perturbations to fluorophore environment. Initially, standard fluorescent dyes were studied and then this FLIM microscope was applied to the imaging of biological tissue, successfully contrasting different tissues and different states of tissue using autofluorescence. To demonstrate the potential for real-world applications, the FLIM microscope has been configured using potentially compact, portable and low cost all-solid-state diode-pumped laser technology. Whole-field FLIM with optical sectioning (3D FLIM) has been realized using a structured illumination technique.     

Spatial calibration of structured illumination fluorescence microscopy using capillary tissue phantoms

Quantitative assessment of microvascular structure is relevant to the investigations of ischemic injury, reparative angiogenesis and tumor revascularization. In light microscopy applications, thick tissue specimens are necessary to characterize microvascular networks; however, thick tissue leads to image distortions due to out-of-focus light. Structured illumination confocal microscopy is an optical sectioning technique that improves contrast and resolution by using a grid pattern to identify the plane-of-focus within the specimen. Because structured illumination can be applied to wide-field (nonscanning) microscopes, the microcirculation can be studied by sequential intravital and confocal microscopy. To assess the application of structured illumination confocal microscopy to microvessel imaging, we studied cell-sized microspheres and fused silica microcapillary tissue phantoms. As expected, structured illumination produced highly accurate images in the lateral (X-Y) plane, but demonstrated a loss of resolution in the Z-Y plane. Because the magnitude of Z-axis distortion was variable in complex tissues, the silica microcapillaries were used as spatial calibration standards. Morphometric parameters, such as shape factor, were used to empirically optimize Z-axis software compression. We conclude that the silica microcapillaries provide a useful tissue phantom for in vitro studies as well as spatial calibration standard for in vivo morphometry of the microcirculation.     

Fluorescence saturation in confocal microscopy

The effects of fluorescence saturation on imaging in confocal microscopy have been studied. To include saturation it was necessary to deviate from the widely assumed linear relationship between the fluorescence and the illumination intensity. The lateral response for a point-like object, as well as the optical sectioning power, decreases depending on the degree of saturation. For very high illumination intensities the response for a saturated point object approached that of a conventional fluorescence microscope in which the fluorescence was not saturated. The decrease in the axial confocal response has been confirmed qualitatively by experiment.     

Two-photon fluorescence surface wave microscopy

This paper demonstrates the principle of two-photon surface wave microscopy with a view to applications on biological samples. We describe a modified scanning optical microscope, which uses specially prepared coverslips. These coverslips are designed to support the propagation of surface waves capable of large field enhancements. We also discuss the beam conditioning necessary to ensure efficient use of the available illumination. Two-photon surface wave fluorescent excitation is demonstrated on fluorescent nanospheres, demonstrating a point spread function width of ≈220 nm at an illumination wavelength of 925 nm. The potential of non-linear surface wave excitation for both fluorescence and harmonic imaging microscopy is discussed.     

Selectable light‐sheet uniformity using tuned axial scanning

Light‐sheet fluorescence microscopy (LSFM) is an optical sectioning technique capable of rapid three‐dimensional (3D) imaging of a wide range of specimens with reduced phototoxicity and superior background rejection. However, traditional light‐sheet generation approaches based on elliptical or circular Gaussian beams suffer an inherent trade‐off between light‐sheet thickness and area over which this thickness is preserved. Recently, an increase in light‐sheet uniformity was demonstrated using rapid biaxial Gaussian beam scanning along the lateral and beam propagation directions. Here we apply a similar scanning concept to an elliptical beam generated by a cylindrical lens. In this case, only z‐scanning of the elliptical beam is required and hence experimental implementation of the setup can be simplified. We introduce a simple dimensionless uniformity statistic to better characterize scanned light‐sheets and experimentally demonstrate custom tailored uniformities up to a factor of 5 higher than those of unscanned elliptical beams. This technique offers a straightforward way to generate and characterize a custom illumination profile that provides enhanced utilization of the detector dynamic range and field of view, opening the door to faster and more efficient 2D and 3D imaging.     

折反式眼底相机光学系统设计

为控制传统眼底相机的杂光和鬼像,设计了一款40°视场、48mm工作距离的折反式眼底相机光学系统。设计了离轴反射式网膜物镜,引入了自由曲面以校正其离轴像差,成像物镜中采用两个自由曲面对网膜物镜的剩余像差进行校正。建立了一种离焦眼模型,用于优化成像光路,消除人眼像差对成像的影响,同时得到不同视度缺陷眼的成像光路。照明光路中使用3个相邻的环形光阑,减少了眼球光学系统反射的杂光。成像光学系统可在-10～+10m-1调焦,物方各视场分辨率为33lp/mm,系统畸变小于8.5%;照明光学系统在不产生鬼像的前提下,可均匀照明眼底,照度非均匀性在15%以内。实验表明,引入自由曲面的折反式眼底相机,有效地消除了杂光和鬼像,满足大视场和大工作距离的要求。     

Combining AFM and FRET for high resolution fluorescence microscopy

Here we demonstrate a new microscopic method that combines atomic force microscopy (AFM) with fluorescence resonance energy transfer (FRET). This method takes advantage of the strong distance dependence in Förster energy transfer between dyes with the appropriate donor/acceptor properties to couple an optical dimension with conventional AFM. This is achieved by attaching an acceptor dye to the end of an AFM tip and exciting a sample bound donor dye through far-field illumination. Energy transfer from the excited donor to the tip immobilized acceptor dye leads to emission in the red whenever there is sufficient overlap between the two dyes. Because of the highly exponential distance dependence in this process, only those dyes located at the apex of the AFM tip, nearest the sample, interact strongly. This limited and highly specific interaction provides a mechanism for obtaining fluorescence contrast with high spatial resolution. Initial results in which 400 nm resolution is obtained through this AFM/FRET imaging technique are reported. Future modifications in the probe design are discussed to further improve both the fluorescence resolution and imaging capabilities of this new technique.