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随着医疗诊断需求的增加,生物分子检测技术越来越受到人们的重视,液相生物芯片技术作为一种高通量,多通道的分子检测手段在近几年得到了飞速发展。通过层层自组装方法制备以微片为载体的拉曼光谱编码液相生物芯片,并利用自行搭建的一套高灵敏度、高分辨率的光学系统,实现对液相生物芯片的定性与定量分析。光学系统由拉曼光谱检测系统与荧光显微成像系统耦合而成。在拉曼光谱检测系统中激光器发射出785 nm波长的激光,通过二向色镜,带反反射镜与物镜汇聚到样品上,样品产生的拉曼散射光,经物镜,带反反射镜,二向色镜与拉曼滤波片,最后通过凹透镜聚焦到光谱仪的狭缝上,光谱仪色散实现在线阵CCD上拉曼光谱的获取。荧光显微成像系统应用光学成像原理,通过调节凹透镜与405 nm的激发光之间的距离,使激发光通过物镜均匀的照射到样品之上,样品激发出的荧光,通过物镜,带反反射镜,二向色镜,滤波片与相应的凹透镜,最后成像到面阵CCD上。改进传统便携式拉曼光谱检测系统光路并选用相应波段的带反反射镜与焦距20倍的物镜完成拉曼光谱检测系统与荧光显微成像系统的耦合。为了减少两路系统之间的相互影响选用合适的二向色镜以及滤波片,在提高耦合系统获取数据的准确性中有着重要的作用。该系统通过对反应之后的液相生物芯片进行拉曼光谱检测,以完成对每个编码玻片的定性识别,即解码;同时激发反应后液相生物芯片的荧光并采集荧光强度图,根据每个解码玻片上的荧光强度值完成对目标检测物的定量分析。区别于传统荧光编码液相生物芯片, 拉曼光谱编码具有稳定性更强,光谱分辨率更高等优点。该光学系统集拉曼光谱检测系统与荧光显微成像系统于一体,解决了目前未有基于拉曼编码的液相生物芯片的检测系统的问题,并且可同时对多种目标物进行识别和定量分析,提升了实验结果的准确性。 相似文献
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利用光锥耦合的ICCD系统探测荧光染料标记的生物芯片,并对CCD芯片和像增强器制冷,以提高探测灵敏度。基于实验分析结果,指出背景噪声的主要来源为杂散光和生物芯片基底所发的荧光,指出用镜头成像限制了系统探测灵敏度的提高,可采用低荧光物质作为生物芯片的基底对系统加以改进。 相似文献
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给出计算机辅助装调中失调量与像差关系的数学模型,利用Zemax对折轴三反射光学系统进行失调仿真,得到失调数据,再对数据进行分析和处理,求得次镜和三镜的灵敏度矩阵,还对各种初级像差与各装调参量之间的对应关系及该系统的失调特性进行讨论。根据以上分析结果,最终确定了共轴三反射光学系统的装调方案。为了验证方案的可行性,在Zemax中对共轴三反射光学系统人为地加入不同的失调量后,按照确定的装调方案,利用建立的计算机辅助装调数学模型计算出失调量的大小和方向,再根据计算结果对共轴三反射光学系统进行调整,仿真结果和实际装调结果都证明该方案是可行的。 相似文献
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为优化毛细管电泳荧光信号检测系统,提高检测灵敏度,以100~1 000碱基对的脱氧核糖核酸作为分离对象,羟乙基纤维素为筛分介质,研究了直流电场下毛细管电泳荧光信号检测系统中的噪声特性.对不同分离电场强度、羟乙基纤维素溶液浓度和分子量、毛细管有效长度以及毛细管内径形状等情况下的噪声特性进行分析.分析得到该检测系统中信噪比最佳的优化参量,即分离电场强度为500~600V/cm、羟乙基纤维素浓度为0.6%~0.7%、羟乙基纤维素分子量为250、圆形内径为50μm以及毛细管有效长度为8cm. 相似文献
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设计了DNA分析仪,采用激光诱导荧光对样品的毛细管电泳结果进行检测,对仪器的灵敏度进行了理论分析和实验测试.分析表明:荧光的探测灵敏度取决于入射到光探测器(PMT或CCD)上的荧光强度及光探测器自身的探测能力.结合设计参量,计算得到DNA分析仪针对染料TO(thiazole orange)作为荧光标记的探测灵敏度为0.127 fluor/μm3个荧光分子.和实验结果相比,双链DNA样品pGEM-3Zf(+)/Hae III Markers加TO进行毛细管电泳实验,得到实际的探测灵敏度为0.145 4 fluor/μm3荧光分子.研究表明:提高探测灵敏度的措施是制备性能良好的毛细管涂层和合理使用筛分介质,合理增大激光功率、尽可能增大物镜的数值孔径、提高光电倍增管的探测下限. 相似文献
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为了提高大靶面交汇测量系统的测量准确度,分析了交汇测量原理,推导出脱靶量坐标公式.根据脱靶量公式分析其各项参量,利用几何关系建立像元坐标与偏移角度之间的映射模型.根据映射模型,利用光栅尺设计了一种针对线阵相机的标定方法,该方法不考虑相机参量,将整个光学系统看作一个整体,基于整体参量直接对像元坐标和它所对应的偏移角进行标定.实验结果表明,标定后的交汇测量系统在1.4m处的平均测量误差为0.4mm,最大测量误差优于0.6mm.该方法简单高效,可提高系统标定的速度,且标定误差满足系统交汇测量准确度的要求. 相似文献
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为分析平顶高斯光束通过光学系统传输时圆孔光阑失调和光学元件失调对平顶高斯光束传输特性的影响,利用失调圆孔光阑的近似展开式和适用于失调光学系统的广义衍射公式,得出了平顶高斯光束经含失调圆孔光阑的失调光学系统传输的近似解析式,给出了输出光束场分布与光束参量、光阑孔径尺寸、光阑和光学元件失调量等的定量关系.针对特定光学系统定量分析了各失调量对输出光束场分布的影响,结果表明各元件失调都对输出光束强度分布产生较大影响.但在各失调量较小的情况下,透镜失调对输出光束传输特性的影响比光阑失调对输出光束传输特性的影响更明显. 相似文献
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Calcein is an intracellular fluorescent probe that has been used as an indicator of cell volume in several previous studies. These studies have reported two different fluorescence responses depending on the optical setup used to collect the data: wide-field microscopy has resulted in a decrease in fluorescence upon cell shrinkage, whereas confocal microscopy has been shown to yield the opposite result. In this short communication, we have investigated the effect of optical setup on detection of cell volume changes in calcein-stained endothelial cells. A confocal microscope was used to collect the fluorescence data, and the pinhole diameter was varied in order to examine the effects of optical section thickness on fluorescence response. For large pinhole diameters – which correspond to relatively thick optical sections – fluorescence intensity decreased when cells were induced to shrink. In contrast, for small pinhole diameters the fluorescence intensity increased with cell shrinkage. The transition between these two types of fluorescence responses occurred when using a pinhole diameter of 285 μm, which corresponds with an optical section thickness slightly less than the height of the cells. Our results have implications for the design and interpretation of experiments involving the use of calcein as a cell volume indicator. 相似文献
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A new three-dimensional (3D) optical fluorescent tomographic imaging scheme is proposed with structured illumination and spatial Fourierdomain decomposition methods for the first time. In this spatial Fourier-decomposition optical fluorescence tomography (SF-OFT), the intensity of focused excitation light from an objective lens is modulated to be a cosine function along the optical axis of the system. For a given position in a two-dimensional (2D) raster scanning process, the spatial frequency of the cosine function along the optical axis sweeps in a proper range while a series of fluorescence intensity are detected accordingly. By making an inverse discrete cosine transformation of these recorded intensity profiles, the distribution of fluorescent markers along the optical axis of a focused laser beam is obtained. A 3D optical fluorescent tomography can be achieved with this proposed SF-OFT technique with a simple 2D raster scanning process. 相似文献
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Martin J. Bishop Gil Bub Alan Garny David J. Gavaghan Blanca Rodriguez 《Physica D: Nonlinear Phenomena》2009,238(11-12):1008-1018
Photon scattering is known to distort the fluorescence signals recorded from optically mapped cardiac tissue. However, the contribution of the parameters which define the optical detection set-up has not been assessed. In this study, Monte Carlo (MC) simulations of photon scattering within ventricular tissue are combined with a detailed model of a tandem-lens optical detection apparatus to characterise (i) the spatial origin upon emission of photons recorded in voltage-sensitive fluorescence measurements of cardiac electrical activity (using the fluorescent dye di-4-ANEPPS) and how this affects signal distortion, and (ii) the role the detector characteristics could play in modulating signal distortion during uniform illumination and photon emission from tissue depth. Results show that, for the particular excitation/emission wavelengths considered (488 nm and 669 nm, respectively), the dimensions of the scattering volume during uniform illumination extend around 3 times further in the surface recording plane than in depth. As a result, fluorescence recordings during electrical propagation are more distorted when transmembrane potential levels differ predominantly in the surface plane than in depth. In addition, MC simulation results show that the spatial accuracy of the fluorescence signal is significantly limited due to photon scattering, with only a small fraction of the recorded signal intensity originating from tissue beneath the pixel (approximately 11% for a 0.25×0.25 mm pixel). Increasing pixel size increases this fraction, however, it also results in an increase in the scattering volume dimensions, thus reducing the spatial resolution of the optical system, and increasing signal distortion. MC simulations also demonstrate that photon scattering in cardiac tissue limits the ability of optical detection system tuning in accurately locating fluorescent emission from depth. Specifically, our results prove that the focal plane depth that yields maximum signal intensity provides an underestimation of the emission depth. In conclusion, our study demonstrates the potential of MC simulations of photon scattering in guiding the design of optical mapping set-ups to optimise performance under diverse experimental conditions. 相似文献
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光纤生物传感器是现代生物传感技术中的一个非常重要的类别。当前,许多以荧光检测为手段的光纤生物传感器已经商品化了,但几乎都是依靠检测荧光指示剂的光强来获取生物信息,而直接利用生物样品的自体荧光光谱来获取生物信息的光纤传感系统却还未上市。利用自行研发的一套三维荧光光谱光纤传感系统对新鲜的人体乳腺组织切片进行了研究,实验结果表明乳腺癌变组织与正常组织的自体荧光光谱的峰位和峰强比值存在明显区别,这是因为癌变组织的生化成分发生了根本改变。尽管其间的规律还需进一步探索,但可以展望,自体荧光光谱技术与光纤生物传感技术的结合有潜力成为人体恶性肿瘤在线原位诊断的有利工具。 相似文献
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The calcium-ion indicator dye, Calcium Green 1 (CG-1), has been characterized using a combination of ensemble and single-molecule
optical spectroscopy measurements. In terms of ensemble measurements, CG-1 demonstrated a strong increase in fluorescence
emission as a function of increasing [Ca2+]. This was accompanied by a change in the relative proportions of two chemical forms of the dye, each with a different fluorescence
lifetime, which were found to co-exist in solution. From single-molecule fluorescence measurements, it was found that the
fluorescence intensity and photobleaching time (on-time) of each CG-1 molecule was invariant with [Ca2+] and that changes in ensemble fluorescence intensity simply correlates with the number of fluorescent molecules in solution.
These results are compared with that of the related system, Calcium Green 2 (CG-2), and the mechanisms of operation of these
two indicator dyes are discussed. 相似文献
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基于激光光源和氙灯光源在线荧光光谱法测定罗丹明B、维生素B2、荧光素和异硫氰酸荧光素含量比较研究激光光源产生的荧光强度和氙灯光源产生的荧光强度。罗丹明B、维生素B2、荧光素和异硫氰酸荧光素浓度均为10 μg·mL-1,积分时间100 ms,测定3次得其平均值。在线荧光光谱法最大吸收波长分别为580,450,488和510 nm;最大发射波长依次为594,530,525和524 nm。紫外-可见分光光度法测得其最大吸收波长为557,441,481和490 nm;荧光分光光度法测得其最大发射波长为586,520,519和520 nm。通过测定药物,发现激光光源产生的荧光光强度较强于氙灯光源产生的荧光光强度,原因不仅跟光源有关,而且与药物分子的共轭体系大小、共轭大π键的共平面性及其刚性程度、分子母体上取代基的种类有关,分子所处的外界环境如温度、溶剂、溶液酸碱度、激发光的照射等因素也会影响荧光效率。激光光源和氙灯光源产生的荧光光强度大小顺序为罗丹明B>荧光素>异硫氰酸荧光素>维生素B2。激光光源在线荧光光谱法在一定程度上填补了在线荧光光谱仪在食品、药品痕量检测方面应用的空白。 相似文献
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针对乙醇水溶液荧光发射的四个特征参量进行了研究,得出了该溶液发射荧光光子的时域和频域特征参量。发射光谱和激发光谱表明乙醇水溶液中含有三个结构不同的发光物质,其发射峰分别位于290 nm,305 nm和330 nm处,与其相对应的最佳吸收峰为265 nm,280 nm和236 nm。荧光强度随溶液中乙醇与水体积混合比的变化规律也证实了三种不同发光结构的存在。在荧光光谱峰值波长处分别监测其荧光强度随时间的衰变过程,将获得的荧光衰减动力学曲线采用指数方法拟合并进行解卷积处理,测试的荧光寿命分别对应8 ns,12 ns,25 ns。结合乙醇水溶液荧光发射的四个特征参量可以看出:乙醇分子和水分子发生团簇作用形成了三个新的分子结构从而可发射具有不同能量的荧光光子。该研究结果能为乙醇水分子的团簇结构研究提供参考。 相似文献