共焦扫描显微成象的光学断层平面分辨率与信噪比之间的关系

在Fried的一维分辨度理论的基础上,系统地讨论了光学成象系统两维分辨率与信噪比之间的定量关系,并以反射式共焦显微成象为例,建立了实际显微成象系统分辨率的定量计算方法.所得出的结果对于选择共焦扫描显微成象系统的最佳参量及评价所设计的显微成象系统的性能具有重要的意义.     

荧光寿命成象显微技术及其应用

本文综述了荧光寿命成象显微技术的概念、原理及实现方法,介绍了荧光寿命成象显微技术在生物物理、生物化学及临床医学诊断等领域的最新研究成果和发展现状,并就其未来的发展及应用研究进行了讨论.     

生物样品的软X射线显微成象

软X射线显微术最适合于自然状态下生物样品的高分辨率成象。在过去十年中软X射线显微成象技术在世界范围内得到了迅速的发展。本文扼要地讨论了软X射线显微成象的衬度机制和高分辨率X射线光学的基本原理,并且综述介绍各种软X射线显微成象方法以及它的发展和前景。     

生物样品的软X射线显微成象

软X射线显微术最适合于自然状态下生物样品的高分辨率成象。在过去十年中软X射线显微成象技术在世界范围内得到了迅速的发展。本文扼要地讨论了软X射线显微成象的衬度机制和高分辨率X射线光学的基本原理,并且综述介绍各种软X射线显微成象方法以及它的发展和前景。     

共焦显微镜三维轮廓快速测量方法及其误差分析

共焦显微术在生物学及工程应用中已成为一种有效的测量、观察方法。本文针对共 焦显微三维测量系统提出了一种高精度快速算法,并进行了误差分析与计算机仿真。对实际 共焦三维测量系统的设计具有重要意义。     

基于共焦原理的反射式自聚焦光纤传感技术研究

基于共焦显微成象原理,提出了一种由自聚焦透镜和X型光纤组成的高精度反射式自聚焦光纤传感器.该传感器具有绝对位置跟踪的特殊能力,并有体积小、抗干扰性强、轴向分辨率高等特点.由实验分析得,其轴向分辨率可达30nm.可用于表面微观形貌、外形尺寸、曲面轮廓等测量领域.     

共焦扫描荧光显微镜的信噪比与测量精度

激光扫描共焦显微术和多光子显微术等新的显微成像技术可以对厚的生物样品实现光学断层成像 ,因而在生物医学诊断领域具有重要的应用前境。在Fried的一维分辨度理论的基础上 ,系统地讨论了运用共焦扫描荧光显微术在进行光学断层成像时 ,其光学断层平面分辨度与信噪比之间的定量关系 ,建立了实际显微成像系统平面测量精度的定量计算方法。所得出的结果对于选择共焦扫描显微成像系统的最佳参数及评价所设计的显微成像系统的性能具有重要的意义。     

共焦扫描显微成象系统的点扩散函数再商讨

针对共焦扫描显微成象中物体相对于被扫描点的位置矢量是反向移动的特性,探讨了物体移动后的物函数在此基础上,根据成象的理论进行了计算,得到新的系统点扩散函数表达式,最后根据计算结果分析成象的性质,得出了系统成象的相干性.     

扫描显微系统中各组元对成像的影响

利用光强点扩散函数讨论扫描显微探测系统中各组无形状对成像的影响。由光源和探测器的讨论得出共焦成像术具有最高横向分辨率的结论，阐明共焦系统中透镜（环瞳和方瞳）对成像的影响，并与圆瞳进行比较，得到最佳系统匹配方法。     

激光分光瞳共焦显微镜系统研制

为了满足生物类等样品对大工作距和高分辨率共焦显微镜的需求,将分光瞳技术与激光共焦显微技术结合应用到成像系统上。阐述了激光分光瞳共焦显微成像原理,首次成功搭建了相应的显微镜成像测量系统。理论分析和实验表明:分光瞳共焦显微技术独特的非共轴结构使系统的轴向分辨力是相同数值孔径物镜单轴系统的3倍以上,对理论高度为100nm的台阶样品进行成像测试,得到的样品三维形貌,成像质量良好。     

Spatial and temporal coherence effects in interference microscopy and full‐field optical coherence tomography

Low‐coherence optical microscopy or optical coherence microscopy uses light with short coherence length. The well‐known case is: “white‐light interferometry”, which became recently more known as: “optical coherence tomography”. However, when lenses and microscope objectives are used to create interferometric images, in what is known classically as “interference microscopy” or today as “full‐field optical coherence tomography” the spatial coherence starts to play a critical role. In this article the coherence effects in low‐coherence optical microscopy are reviewed. As this technology is becoming increasingly publicized due to its importance in three‐dimensional imaging, particularly of scattering biological media and optical metrology, the understanding of the fundamental physics behind it is essential. The interplay between longitudinal spatial coherence and temporal coherence and the effects associated with them are discussed in detail particularly when high numerical apertures are used. An important conclusion of this study is that a high‐contrast, high‐resolution system for imaging of multilayered samples is the one that uses narrowband illumination and high‐NA objectives with an index‐matching fluid. Such a system, when combined with frequency‐domain operation, can reveal nearly real‐time three‐dimensional images, and is thus competitive with confocal microscopy.     

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.     

Engineered microsphere contrast agents for optical coherence tomography

Contrast agents are utilized in virtually every imaging modality to enhance diagnostic capabilities. We introduce a novel class of optical contrast agent, namely, encapsulating microspheres, that are based not on fluorescence but on scattering nanoparticles within the shell or core. The agents are suitable for reflection- or scattering-based techniques such as optical coherence tomography, light microscopy, and reflectance confocal microscopy. We characterize the optical properties of gold-, melanin-, and carbon-shelled contrast agents and demonstrate enhancement of optical coherence tomography imaging after intravenous injection of such an agent into a mouse.     

生物医学领域中超连续激光光谱技术的研究进展

超连续谱激光指的是当泵浦激光穿过特殊光波导时,一系列的非线性效应引起入射激光束的光谱展宽,从而输出宽光谱激光束。随着超快激光和光子晶体光纤技术的发展,利用超短脉冲在光子晶体光纤中的传播链产生相干的且亮度高的超连续谱激光成为了一种理想的白光源。自从超连续谱激光源投入应用以来,其应用领域越来越广。尤其在生物医学的细胞、血液等样品分析当中,荧光光谱学、流式细胞仪、共焦显微、光学相干层析等技术都是强有力的分析工具,在采用这些先进技术的科学仪器中,超连续谱激光源成为了一种主要光学部件。首先对超连续谱激光源的国际研究进展作了详细介绍,然后对超连续激光光谱技术在显微成像、流式细胞仪、荧光寿命成像显微、荧光共振能量转移、光学相干层析、共焦显微生物医学分析等生物医学领域中的发展及应用作了综合阐述。对超连续激光光谱技术在非接触式血液制品鉴别的需求、方案及研究进展进行了重点论述,包括覆盖400～2 000 nm光谱范围的光纤化轻型超连续谱激光光源研究;采用超连续谱激光光谱方法探索不同物种血液的种属特征;根据大数据的血液样品光谱特征元数据库分析建立数学模型,利用数学模型实现对血液样品种属光谱学判定;血液鉴别光谱分析便携式整机系统研发等。对超连续激光光谱技术在生物医学领域的应用前景作了展望。     

Microstructure measurement based on frequency-shift feedback in a-cut Nd:YVO_4 laser

A new optical method based on frequency-shift feedback and laser confocal microscopy is presented to noninvasively measure a microstructure inside a sample. Due to the limit of axial resolution caused by poor signal detection ability, conventional laser feedback cannot precisely measure the microstructure. In this Letter, the light scattered by the sample is frequency shifted before feedback to the laser to obtain a magnification. Weak signals that change with the microstructure can be detected. Together with the tomography ability of laser confocal microscopy, the inner microstructure can be measured with high axial resolution.     

Numerical correction of coherence gate in full-field swept-source interference microscopy

A big problem in low-coherence interference microscopy is the degradation of the coherence signal caused by shift of the angular and temporal spectrum gates. It limits the depth of field in confocal optical coherence microscopy and degrades images of sample inner structure in most interference microscopy techniques. To overcome this problem we propose numerical correction of the coherence gate in application to full-field swept-source interference microscopy. The proposed technique allows three-dimensional sample imaging without mechanical movement of the microscope components and is also capable of determining separately the geometrical thickness and the refractive index of the sample layers, when the sample contains a transversal pattern. The applicability of the proposed technique is verified with numerical simulation.     

Photoacoustic microscopy

Photoacoustic microscopy (PAM) is a hybrid in vivo imaging technique that acoustically detects optical contrast via the photoacoustic effect. Unlike pure optical microscopic techniques, PAM takes advantage of the weak acoustic scattering in tissue and thus breaks through the optical diffusion limit (∼1 mm in soft tissue). With its excellent scalability, PAM can provide high‐resolution images at desired maximum imaging depths up to a few millimeters. Compared with backscattering‐based confocal microscopy and optical coherence tomography, PAM provides absorption contrast instead of scattering contrast. Furthermore, PAM can image more molecules, endogenous or exogenous, at their absorbing wavelengths than fluorescence‐based methods, such as wide‐field, confocal, and multi‐photon microscopy. Most importantly, PAM can simultaneously image anatomical, functional, molecular, flow dynamic and metabolic contrasts in vivo. Focusing on state‐of‐the‐art developments in PAM, this Review discusses the key features of PAM implementations and their applications in biomedical studies.     

Hybrid Rayleigh,Raman and two‐photon excited fluorescence spectral confocal microscopy of living cells

A hybrid fluorescence–Raman confocal microscopy platform is presented, which integrates low‐wavenumber‐resolution Raman imaging, Rayleigh scatter imaging and two‐photon fluorescence (TPE) spectral imaging, fast ‘amplitude‐only’ TPE‐fluorescence imaging and high‐spectral‐resolution Raman imaging. This multi‐dimensional fluorescence–Raman microscopy platform enables rapid imaging along the fluorescence emission and/or Rayleigh scatter dimensions. It is shown that optical contrast in these images can be used to select an area of interest prior to subsequent investigation with high spatially and spectrally resolved Raman imaging. This new microscopy platform combines the strengths of Raman ‘chemical’ imaging with light scattering microscopy and fluorescence microscopy and provides new modes of correlative light microscopy. Simultaneous acquisition of TPE hyperspectral fluorescence imaging and Raman imaging illustrates spatial relationships of fluorophores, water, lipid and protein in cells. The fluorescence–Raman microscope is demonstrated in an application to living human bone marrow stromal stem cells. Copyright © 2009 John Wiley & Sons, Ltd.     

微型差动式共焦自聚焦光聚焦探测系统

为解决微小内轮廓尺寸为代表的微小尺寸的非接触式超精密测量问题，提出了将自聚焦透镜体积小的特点与共焦显微技术的高分辨率和绝对位置跟踪特性相结合的差动式自聚焦共焦微型显微技术的光探测技术，建立了相应的传感系统，介绍了系统的工作原理和构成，自聚焦透镜测头直径为1mm，两个探测器差动设置，不但消除了光源的光强漂移和探测器的电子漂移产生的共模噪声，提高了测量信噪比，而且有效地提高了系统的轴向分辨率，初步实验表明，系统轴向分辨率在倾斜率小于20度的范围内可达5nm。     

Fiber-optic-bundle-based optical coherence tomography

A fiber-optic-bundle-based optical coherence tomography (OCT) probe method is presented. The experimental results demonstrate this multimode optical fiber-bundle-based OCT system can achieve a lateral resolution of 12 microm and an axial resolution of 10 microm with a superluminescent diode source. This novel OCT imaging approach eliminates any moving parts in the probe and has a primary advantage for use in extremely compact and safe OCT endoscopes for imaging internal organs and great potential to be combined with confocal endoscopic microscopy.