In vivo confocal imaging of fast intrinsic optical signals correlated with frog retinal activation

Using freshly isolated animal retinas, we have conducted a series of experiments to test fast intrinsic optical signals (IOSs) that have time courses comparable to electrophysiological kinetics. In this Letter, we demonstrate the feasibility of in vivo imaging of fast IOSs in intact frogs. A rapid line-scan confocal ophthalmoscope was constructed to achieve high-speed IOS recording. By rejecting out-of-focus background light, the line-scan confocal imager provided the resolution to differentiate individual photoreceptors in vivo. Rapid confocal imaging disclosed robust IOSs with time courses comparable to retinal electroretinogram kinetics. High-resolution IOS images revealed both positive (increasing) and negative (decreasing) light responses, with subcellular complexity.     

Large area confocal microscopy

Imaging large tissue areas with microscopic resolution in vivo may offer an alternative to random excisional biopsy. We present an approach for performing confocal imaging of large tissue surface areas using spectrally encoded confocal microscopy (SECM). We demonstrate a single-optical-fiber SECM apparatus, designed for imaging luminal organs, that is capable of imaging with a transverse resolution of 2.1 microm over a subsurface area of 16 cm2 in less than 1 min. Due to the unique probe configuration and scanning geometry, the speed and resolution of this new imaging technology are sufficient for comprehensively imaging large tissues areas at a microscopic scale in times that are appropriate for clinical use.     

Measuring tablet porosity using multispectral imaging system

A multispectral imaging system with computer controlled light source of 16 light emitting diodes is applied for measuring of tablet porosity. The system is based on a subspace vector model of surface reflection. The measured spectral data are compressed on the stage of measurement and used directly for the discrimination of tablets with different porosity. The experimental results demonstrate that the multispectral imaging system is a potential method for tablet porosity measurement.     

Developing a contact probe for rodent fundus imaging in a confocal scanning laser ophthalmoscope

Since significant ocular differences in both anatomical structure and optical properties exist between rodents and humans, clinical imaging devices for human use are not suitable for use on rodents. In this study, we develop a contact probe with a flexible surface that can closely fit the rodent cornea for fundus imaging with a confocal scanning laser ophthalmoscope. Both Zemax simulation and in vivo fundus imaging demonstrate that this contact probe can significantly improve both the imaging quality and the operational convenience.     

基于成像系统物理特性的多光谱图像与全色波段图像融合

提出了一种基于成像系统物理特性的多光谱图像与全色波段图像融合算法。该算法采用àtrous小波变换提取全色波段图像的空间细节信息,并将提取的空间信息按照一定的注入模型调整后添加到各波段多光谱图像中去,得到具有高空间分辨力的多光谱图像。注入模型充分考虑了各波段成像传感器的相对光谱响应函数、地表物体对各波段的光谱反射率以及各波段的辐射调整系数等成像系统的物理特性,使融合后的多光谱图像在显著提高空间质量的同时,最大可能地保留了原始多光谱图像的光谱特性。对IKONOS卫星遥感影像的融合实验结果表明,该算法在光谱保留和空间质量提高方面较其它基于小波变换的融合算法都具有更高的性能。     

High-resolution optical coherence microscopy for high-speed, in vivo cellular imaging

Optical coherence microscopy (OCM) is demonstrated with a high-speed, broadband, reflective-grating phase modulator and a femtosecond Ti:Al2O3 laser. The novel system design permits high-resolution OCM imaging in a new operating regime in which a short coherence gate is used to relax the requirement for high-numerical-aperture confocal axial sectioning. In vivo cellular imaging is demonstrated in the Xenopus laevis tadpole and in human skin with a 3-microm coherence gate and a 30-microm confocal gate. The ability to achieve cellular imaging with a lower numerical aperture should facilitate the development of miniaturized probes for in vivo imaging applications.     

Multispectral photoacoustic imaging of fluorochromes in small animals

Fluorochromes have become essential reporter molecules in biological research. We show that the depth-resolved distribution of fluorochromes in small animals can be imaged with 25 fmol sensitivity and 150 microm spatial resolution by means of multispectral photoacoustic imaging. The major advantage of the multispectral approach is the sensitive differentiation of chromophores and fluorochromes of interest based on self-reference measurements, as evidenced in this study by resolving a commonly used fluorochrome (Alexa Fluor 750) in mouse. The suggested method is well suited for enhancing visualization of functional and molecular information in vivo and longitudinally.     

Multispectral imaging using multiple-bandpass filters

We present a method for multispectral imaging. This method uses color CCD cameras with a multiple-bandpass filter, which modifies the spectral response of the cameras used and enables concurrent acquisition of multiple images at defined spectral bands. We experimentally demonstrate methodological feasibility using two color CCD cameras and a polychroic mirror to simultaneously capture eight spectral bands. We discuss how the method developed is well suited for multispectral applications of transient phenomena or for real-time measurements.     

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.     

一种可实时化的多光谱图像融合系统

在多光谱摄影和光电成像技术基础上，本文提出了一种可实时化的多光谱图像融合系统。与多光谱摄影相比，不仅可实现多光谱实时化摄像，而且光谱响应进一步向近红外延伸，并引入图像处理技术。实验表明：这种技术对于增加系统的识别能力具有明显的作用，可进一步构成不同用途的新型多光谱图像融合系统。     

基于非采样Contourlet变换的遥感图像融合算法

为了使融合后的多光谱图像在尽可能保持原始多光谱图像光谱特性的同时,显著提高空间分辨力,提出了一种基于非采样Contourlet变换(NSCT)的遥感图像融合算法。算法首先对全色波段图像进行非采样Contourlet变换,得到全色波段图像的低频子带系数和各带通方向子带系数;然后针对多光谱图像的每一个波段,将其进行双线性插值后作为融合后多光谱图像的低频子带系数,对全色波段图像的各带通方向子带系数采用基于成像系统物理特性的注入模型(调整系数)进行局部调整后,作为融合后多光谱图像的各带通方向子带系数,从而得到融合后多光谱图像的非采样Contourlet变换系数;最后再经非采样Contourlet逆变换得到该波段具有高空间分辨力的多光谱图像。采用IKONOS卫星遥感图像进行了仿真实验,实验结果表明,该算法在光谱保留和空间质量提高方面优于其它传统的遥感图像融合算法。     

基于多光谱遥感成像链模型的系统信噪比分析

信噪比(SNR)是评价多光谱遥感成像性能的重要指标,在设计多光谱遥感成像仪的最初阶段应进行分析,从而确定各分系统相关参数。多光谱遥感系统的成像链模型综合考虑辐射源、地物光谱反射、大气辐射传输、光学系统成像、分光元件特性、探测器光谱响应和相机噪声等各个环节,可用于进行成像过程端对端的完整分析。以基于滤光片阵列的多光谱遥感系统为例,采用MODTRAN软件进行大气辐射传输计算,对不同太阳天顶角下,不同目标地物计算像面的照度,根据电荷耦合器件探测器的噪声模型,计算出不同工作条件下多光谱遥感系统的SNR。通过对SNR的分析,可给出该类型多光谱遥感系统获得最佳性能的工作条件,并能够结合使用要求进行光学系统参数的优化选择。     

Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging

We demonstrate a confocal optical microscope that combines cw two-photon-excited fluorescence microscopy with confocal Raman microscopy. With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous Raman imaging. The distribution of the UV-absorbing fluorophore Hoechst 33342 in the apoptotic HeLa cells is measured in the combined cw two-photon-excited fluorescence and Raman microscopy modes. The 647-nm line of a Kr-ion laser is used to excite both the Raman scattering and the two-photon-excited fluorescence emission. The lateral and axial resolutions in the two imaging modes are compared by use of the Gaussian beam approximation and backprojection of the focal volume through the confocal pinhole.     

敦煌壁画绘画材料多光谱图像标准数据库的建立和应用

多光谱摄影图像调查技术,基于不同物质对不同光谱吸收与反射的差异及不同波段单色光谱有相应的灰度偏向,以单色光谱灰度图像经伪彩色强化,可作为一种绘画材料普查方法。这种方法的应用首先需要建立相关遗址绘画材料多光谱图像标准,以此为基础才可以通过多光谱伪彩色等图像对壁画中漫漶壁画、相近色相颜料及无法观察的有机材料做前期普查,为后期高精度仪器的进一步调查提供参考。采用已知敦煌壁画中常用29种绘画材料制作标准颜料色板,通过规范多光谱拍摄程序和优化多光谱拍摄条件,在250～1 300 nm之间获取各种颜料在不同光谱区域的反射及荧光图像,再经过后期科学的图像校正及红外、紫外反射伪彩色图像合成,初步建立敦煌壁画颜料多光谱图像标准数据库。对真实壁画进行多光谱拍摄,将获取的可见反射、红外反射伪彩色和紫外反射伪彩色图像与标准数据库图像进行交叉对比,并采用便携式显微镜、便携式X荧光、近红外光谱等分析技术验证所获取的多光谱分析结果,表明壁画颜料多光谱图像标准数据库的建立,作为一种新型无损检测辅助方法,可通过多光谱图像对敦煌壁画颜料前期鉴别、推断绘画材料属性以及获取颜料分布状况等信息。绘画材料光谱图像数据库的建立可提高常见绘画材料类别及范围的前期调查效率和可靠性,是一种有效的颜料面集调查辅助方法。     

高速多光谱 TDI CCD 成像电路系统

为实现多光谱TDI CCD的高速高信噪比成像,利用可空间应用的多光谱TDI CCD传感器研制出了高性能成像电路系统。该系统以现场可编程门阵列（ FPGA）为核心逻辑单元,带有RS422外围通信控制接口,并采用CAMERALINK接口输出图像数据。系统具有动态推扫成像的能力,可同时输出全色和彩色两种模式的图像数据。利用灰度条纹的靶标对传感器的3个多光谱（ R、G、B）感光区标定白平衡,利用彩色条纹的靶标对系统进行成像测试,在驱动频率为15 MHz的情况下,系统单片CCD输出的图像数据率达到1.2 Gbps。试验结果表明,获取全色图像的信噪比达到了53.56 dB,各多光谱图像的信噪比较高的也在40 dB以上,满足空间对地高分辨多光谱遥感成像的技术指标要求,对高速空间多光谱遥感相机的研制具有借鉴意义。     

多光谱荧光图像技术检测农药残留

多光谱成像技术是一种结合传统的二维成像技术及光谱技术的新兴无损检测技术。本文利用有机磷农药的荧光特性,结合多光谱成像技术和荧光激发技术,研发了叶片农药残留快速检测的多光谱荧光成像系统。结果表明,在190~300 nm的紫外光激励下,配置好的有机磷农药氧乐果会在440 nm附近产生显著的荧光发射,并且荧光发射光谱峰值与农药浓度具有良好的线性关系。滤光片可置换的多光谱成像系统研发以及实验检测结果为开发实时在线的农药残留快速无损检测系统提供了技术保障和理论依据。     

Multi-color miniature dual-axis confocal microscope for point-of-care pathology

We present a miniature microelectromechanical systems-based dual-axis confocal microscope capable of spatially coregistered fluorescence and reflectance imaging at multiple wavelengths. This device has a 10 mm diameter scan head with a 2 mm diameter tip for convenient use during surgery to guide tumor resection. The microscope has an adjustable focal depth of 20-200 micrometers and is capable of imaging with an axial resolution of 9 micrometers and in-plane resolution of 4 micrometers over a field of view of 450×450 micrometers. Simultaneous two-color imaging of individual optical sections is achieved by using a pair of grating-prism assemblies to compensate for chromatic dispersion in the 2 mm diameter gradient index relay lens at the distal tip of the device. Experimental measurements of the axial response of the microscope, as well as two-color images of a reflective bar target and fresh mouse brain tissues, demonstrate the performance of our device and its potential for multicolor in vivo optical sectioning microscopy.     

Three-dimensional integral imaging of micro-objects

We propose a method for displaying micro-objects in space that is based on three-dimensional (3D) integral imaging, in which elemental images are calculated from a two-dimensional sampling of the optical field along different depths by use of confocal scanning microscopy. Experimental results are presented to demonstrate that a uniformly magnified 3D biological specimen can be displayed in space, and thus integral imaging can be used for 3D display of confocal microscopy. To the best of our knowledge, this is the first report of 3D integral imaging of (semitransparent) micro-objects.     

Whole-field five-dimensional fluorescence microscopy combining lifetime and spectral resolution with optical sectioning

We report a novel whole-field three-dimensional fluorescence lifetime imaging microscope that incoporates multispectral imaging to provide five-dimensional (5-D) fluorescence microscopy. This instrument, which can acquire a 5-D data set in less than a minute, is based on potentially compact and inexpensive diode-pumped solid-state laser technology. We demonstrate that spectral discrimination as well as optical sectioning minimize artifacts in lifetime determination and illustrate how spectral discrimination improves the lifetime contrast of biological tissue.     

Speckle variance detection of microvasculature using swept-source optical coherence tomography

We report on imaging of microcirculation by calculating the speckle variance of optical coherence tomography (OCT) structural images acquired using a Fourier domain mode-locked swept-wavelength laser. The algorithm calculates interframe speckle variance in two-dimensional and three-dimensional OCT data sets and shows little dependence to the Doppler angle ranging from 75 degrees to 90 degrees . We demonstrate in vivo detection of blood flow in vessels as small as 25 microm in diameter in a dorsal skinfold window chamber model with direct comparison with intravital fluorescence confocal microscopy. This technique can visualize vessel-size-dependent vascular shutdown and transient vascular occlusion during Visudyne photodynamic therapy and may provide opportunities for studying therapeutic effects of antivascular treatments without on exogenous contrast agent.