Excitation-resolved hyperspectral fluorescence lifetime imaging using a UV-extended supercontinuum source

We present a time-gated, optically sectioned, hyperspectral fluorescence lifetime imaging (FLIM) microscope incorporating a tunable supercontinuum excitation source extending into the UV. The system is capable of resolving the excitation spectrum, emission spectrum, and fluorescence decays in an optically sectioned image.     

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.     

Optically sectioned fluorescence lifetime imaging using a Nipkow disk microscope and a tunable ultrafast continuum excitation source

We demonstrate an optically sectioned fluorescence lifetime imaging microscope with a wide-field detector, using a convenient, continuously tunable (435-1150 nm) ultrafast source for fluorescence imaging applications that is derived from a visible supercontinuum generated in a microstructured fiber.     

无扫描光源调制层析成象显微镜(1)-理论

分析普通显微镜的成象过程可以看到,它不能进行层析成象的原因在于并非所有的空间频率都随着离焦量的增加而衰减.通过改装普通显微镜的照明光路,采用调制光源照明物体,从而获得物体的层析象与普通显微镜成象的叠加象,该叠加象经过快速算法运算后,可实时获得类似于共焦显微镜的层析象,最终实现层析成象.本文通过对本显微镜成象光学系统物理模型的简化,解释了该系统的层析能力和解码方法.根据分析可以发现,这种照明调制显微镜实际上是一种选择性照明显微镜,即选择有限宽度的层进行照明.     

Image formation in structured illumination wide-field fluorescence microscopy

We present a theoretical analysis of the image formation in structured illumination wide-field fluorescence microscopy (SIWFFM). We show that the optically sectioned images obtained with this approach possess the optical sectioning strengths comparable to those obtained with the confocal microscope. We further show that the transfer function behaviour is directly comparable to that of the true confocal instrument. The theoretical considerations are compared with and confirmed by experimental results.     

散斑照明宽场荧光层析显微成像技术研究

介绍了一种新的宽场荧光层析显微方法.在传统宽场显微镜中引入散斑图案照明样品,控制散斑图案的动态变化,利用CCD相机记录对应的一系列荧光图像.由于焦平面内强度变化远比焦平面外强度变化剧烈,通过合适的算法能够获得焦平面的层析分辨的荧光显微图像.标定了系统参数,并研究了不同的图像重建算法对系统性能的影响,获得了不同生物组织样品的层析图像.实验表明,该显微方法能用于组织光学切片成像,在临床医学中具有实际应用价值. 关键词： 荧光 散斑照明 荧光显微 层析     

Image formation in conventional brightfield reflection microscopes with optical sectioning property via structured illumination

We present a theoretical analysis of the image formation in conventional brightfield reflection microscopes with optical sectioning property via structured illumination. We show that the optically sectioned images obtained with this approach possess the optical sectioning strengths comparable to those obtained with the confocal microscope. We further show that the transfer function behaviour is directly comparable to that of the true confocal instrument. Some effects of spherical aberrations are included in the analysis. The theoretical considerations are compared with and confirmed by experimental results.     

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.     

Simultaneous optically sectioned fluorescence and optical coherence microscopy with full-field illumination

Full-field optical coherence microscopy (FF-OCM) and optically sectioned fluorescence microscopy are two imaging techniques that are implemented here in a novel dual modality instrument. The two imaging modalities use a broad field illumination to acquire the entire field of view without raster scanning. Optical sectioning is achieved in both imaging modalities owing to the coherence gating property of light for FF-OCM, and a structured illumination setup for fluorescence microscopy. Complementary image data are provided by the dual modality instrument in the context of biological tissue screening. FF-OCM imaging modality shows the tissue microarchitecture, while fluorescence microscopy highlights specific tissue features with cellular-level resolution by using targeting contrast agents. Complementary tissue morphology and biochemical features could potentially improve the understanding of cellular functions and disease diagnosis.     

Method of obtaining optical sectioning by using structured light in a conventional microscope

We describe a simple method of obtaining optical sectioning in a conventional wide-field microscope by projecting a single-spatial-frequency grid pattern onto the object. Images taken at three spatial positions of the grid are processed in real time to produce optically sectioned images that are substantially similar to those obtained with confocal microscopes.     

Holographic data storage with arbitrarily misaligned data pages

An improved postprocessing algorithm that can compensate for arbitrary misregistrations between a detector array and the coherent image of a pixelated two-dimensional data page is described. Previously [Opt. Lett. 26, 542 (2001)], an algorithm was reported in which both linear and quadratic interpixel cross-talk contributions are reallocated to the appropriate neighboring pixels. However, page misalignments close to +/-0.5 pixels could not be corrected to an acceptable bit-error rate because of propagation in the iterative procedure. An improved algorithm is reported in which an intentional magnification error is introduced optically and then corrected during postprocessing. Experimental results from a pixel-matched megapel volume holographic system are presented, showing that the dependence of bit-error rate on transverse detector alignment is entirely removed. This improved procedure can completely bypass constraints on page registration, optical distortion, and material shrinkage that currently hamper page-oriented holographic storage systems.     

结合基于梯度的振铃评价算法的总变分最小化图像分块复原法

为了消除退化函数随空间变化发生变化模糊图像分块复原法子块之间的不平滑拼接缝,提出了一种结合了基于梯度的振铃评价算法梯度振铃评价(GRM)的总变分(TV)最小化分块复原法.根据图像分布及退化类型将模糊图像划分为矩形、环形或其他形状的子块,图像子块之间要留有一定的重叠区;然后对每一个图像子块进行复原,GRM方法是基于图像梯度结构相似度的图像质量评价算法,以GRM作为TV复原算法迭代过程中的收敛条件,可以更好地控制复原图像的振铃;最后去除复原图像子块含振铃波纹的重叠区,拼接得到完整图像.并以矩形分块及环形分块为例,证明该方法可以很好地抑制图像边界振铃效应,克服分块复原法本身的缺陷,得到拼接平滑的完整图像.     

Iterative reconstruction algorithm for optoacoustic imaging

Optoacoustic imaging is based on the generation of thermoelastic stress waves by heating an object in an optically heterogeneous medium with a short laser pulse. The stress waves contain information about the distribution of structures with preferential optical absorption. Detection of the waves with an array of broadband ultrasound detectors at the surface of the medium and applying a backprojection algorithm is used to create a map of absorbed energy inside the medium. With conventional reconstruction methods a large number of detector elements and filtering of the signals are necessary to reduce backprojection artifacts. As an alternative this study proposes an iterative procedure. The algorithm is designed to minimize the error between measured signals and signals calculated from the reconstructed image. In experiments using broadband optical ultrasound detectors and in simulations the algorithm was used to obtain three-dimensional images of multiple optoacoustic sources. With signals from a planar array of 3x3 detector elements a significant improvement was observed after about 10 iterations compared to the simple radial backprojection. Compared to conventional methods using filtered backprojection, the iterative method is computationally more intensive but requires less time and instrumentation for signal acquisition.     

Small-size spectrometer for emission analysis of low-temperature plasma flows

Calculation data for characteristics of concave holographic arrays are reported, allowing development of low-size spectrometers with medium and high spectral resolution. The holographic arrays were fabricated around new subnanostructured films of chalcogenide semiconductors. A medium-resolution, small-size spectrometer was used to measure the distribution of temperature in the gas flow emanating from a DC plasmatorch with sectioned inter-electrode insert by the relative-intensity method. Presently available methods for measuring the temperature of non-uniform optically thin plasma were employed. A comparison of spectral and thermophysical methods for temperature measurements is given.     

平场复用多焦点结构光照明超分辨显微成像

多焦点结构光照明显微技术(multifocal structured illumination microscopy, MSIM)能在50μm的成像深度内和1Hz的成像速度下实现两倍于衍射极限分辨率的提升,相比传统的宽场结构光照明显微技术,具有较大的成像深度和层析能力,更适合应用于厚样品的长时程三维超分辨成像.然而, MSIM存在成像速度慢、图像处理过程复杂等问题.本文提出了一种基于平场复用多焦点结构光照明的快速超分辨显微成像方法和系统(flat-field multiplexed MSIM, FM-MSIM),通过在照明光路中插入光束整形器件,将高斯光束转变为均为分布的平顶光束,提高激发点阵的强度均匀性和扩大视场;通过将每个衍射受限的激发点沿y方向延长,形成新的多路复用多焦点阵照明图案,提高能量利用率,减少扫描步数,进而提高成像速度和信噪比;结合基于多重测量矢量模型的稀疏贝叶斯学习图像重构算法,简化图像重构步骤,在保证空间分辨率的同时实现至少4倍于传统MSIM的成像速度.在此基础上,利用搭建的FM-MSIM系统进行了BSC细胞微管样片和小鼠肾切片标准样片的超分辨成像实验,实验结果证明...     

Optical sectioning using single‐plane‐illumination Raman imaging

Confocal Raman imaging is widely used for optical sectioning of materials. However, for biological applications it often suffers from poor axial resolution, photodamage to the sample of interest, substrate interference, and long acquisition times. We have applied the principles of light sheet microscopy to Raman imaging and show for the first time that optically sectioned Raman images can be obtained in significantly lower acquisition times. Copyright © 2010 John Wiley & Sons, Ltd.     

Surface-reflection elimination in polarization imaging of superficial tissue

A major drawback in polarization gating of light backscattered from tissue is that surface reflections dominate the image. An optically flat plate and matching fluid applied to the tissue surface, combined with off-axis detection, were previously used to address this problem. This approach is often inappropriate or inconvenient for practical use and more importantly can affect the tissue's optical properties. A method is demonstrated that combines images obtained with linearly and circularly polarized light to produce a polarization-gated image that is free from surface reflections and does not require optically flat plates or matching fluid.     

多元探测器在玫瑰扫描红外亚成像系统中的应用

玫瑰扫描红外亚成像系统是利用玫瑰扫描机构和红外单元探测器构成的一种亚成像系统。提出了一种利用多元小面阵来替代单元探测器的红外亚成像系统 ,可以提高成像系统的空间分辨率和帧频。分析了系统的原理 ,并给出了系统的构成框图。此外 ,就玫瑰扫描成像系统中应用多元探测器可能出现的问题进行了讨论 ,并提出了解决方案。结果表明 ,这种多元小面阵玫瑰扫描成像系统的成本远低于焦平面阵列探测器红外成像系统 ,在性能改善后是诸如末制导炮弹等低成本应用场合的理想红外成像系统     

射线检测数字实时成像的不一致性研究

在基于PaxScan4030平板探测器技术的射线检测中,输出图像的像质除受系统噪声的影响外,同时还受探测器固有的光电响应不一致性的影响。通过对影响平板探测器输出图像质量的分析,建立了探测器响应不一致性校正模型。在用常规叠加方法去除噪声的同时,利用校正因子对采集图像进行实时校正,以改善系统性能,提高成像质量。通过实验数据分析和对校正前后输出图像的比较,验证了此方法的可行性,从而为射线检测提供了一种有效的校正方法。