共查询到18条相似文献,搜索用时 140 毫秒
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激光共焦扫描显微镜的分辨率及像差补偿 总被引:1,自引:0,他引:1
从激光共焦扫描显微镜的基本原理出发,深入讨论激光共焦扫描显微镜的深度分辨率和横向分辨率,同时研究三维成像的像差补偿问题。 相似文献
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激光差动共焦显微镜具备高空间分辨率特点,但因其逐点扫描成像方式,扫描时间长,易受三维扫描系统不稳定和环境干扰等影响,产生系统漂移,影响仪器的空间分辨率。利用楔块机构高稳定特点,结合刹车机构的自由抱闸特性,设计了一种新型的轴向升降机构,由此构建了结构更具稳定特性的电动三维扫描系统。稳定性实验验证在搭建的激光差动共焦显微镜上进行,经过监测系统在90min内的轴向位置,轴向漂移小于50nm,与原三维扫描系统漂移140nm对比,漂移速度明显减慢,稳定性有显著提升,进而明显改善了差动共焦显微成像效果。 相似文献
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双光子成像(Two-Photon Imaging)技术以其优越特性被广泛用于活细胞动态三维成像,但光功率极高的短脉冲光对焦平面荧光分子严重的光漂白极大地影响了双光子长时间成像的图像质量,针对双光子荧光漂白问题,本文提出一种优化光照的双光子(Optimized Lighting-Two Photon,OL-TP)成像技术。通过预扫描获取双光子图像分析高低阈值,以预设的高低阈值为标准优化一幅图像中不同区域的光照时长,利用扫描过程中记录的荧光信息和光照时间信息可以重建OL-TP图像,既保证信噪比又降低荧光漂白。重建的OL-TP图像与传统双光子图像基本一致,信噪比略有降低,但图像并未失真。对110 nm的荧光小球样本分别连续取30幅普通双光子和优化光照的双光子图像,到第30幅图时,重建后的优化光照双光子图像比普通双光子图像荧光漂白降低了28.86%。OL-TP通过优化光照时间大幅降低双光子成像的荧光漂白,使双光子荧光显微镜能够更好地对生物样本进行长时间观测。 相似文献
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Theoretical investigation on Raman induced kerr effect spectroscopy in nonlinear confocal microscopy
The imaging theory of Raman induced Kerr effect spectroscopy (RIKES) in nonlinear confocal microscopy is presented in this
paper. Three-dimensional point spread function (3D-PSF) of RIKES nonlinear confocal microscopy in isotropic media is derived
with Fourier imaging theory and RIKES theory. The impact of nonlinear property of RIKES on the spatial resolution and imaging
properties of confocal microscopy have been analyzed in detail. It is proved that RIKES nonlinear confocal microscopy can
simultaneously provide more information than two-photon confocal microscopy concerning molecular vibration mode, vibration
orientation and optically induced molecular reorientation, etc. It is shown that RIKES nonlinear confocal microscopy significantly
enhances the spatial resolution and imaging quality of confocal microscopy and achieves much higher resolution than that of
two-photon confocal microscopy. 相似文献
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TANG Zhilie XING Da & LIU Songhao . Department of Physics South China Normal University Guangzhou China . School for Information Optoelectronic Science Engineering South China Normal University Guangzhou China Correspondence should be addressed to Tang Zhilie 《中国科学G辑(英文版)》2004,47(1):8-16
ItisreportedrecentlythatnonlinearopticalphenomenonofSHGandTHGhasbeenobservedinmanybiologicaltissues[16].SHGandTHGhavebeenusedtoperformthethree-dimensionalimaginginlivingtissuesandhaveattractedmuchattentionrecently.TherearemanyadvantagesofusingSHGandTHGtoperformthethree-dimensionalimaginginlivingtissues,suchasnoninvasiveandnophotobleaching,inadditiontotheimagingpropertiesofmulti-photonfluorescenceimaging[7—9].Firstly,unlikeinthesingle-andmulti-photonfluorescenceprocesses,onlyvirtualstat… 相似文献
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Nicolas Sandeau 《Optics Communications》2006,264(1):123-129
The main advantage of two-photon fluorescence confocal microscopy is the low absorption obtained with live tissues at the wavelengths of operation. However, the resolution of two-photon fluorescence confocal microscopes is lower than in the case of one-photon excitation. The 4Pi microscope type C working in two-photon regime, in which the excitation beams are coherently superimposed and, simultaneously, the emitted beams are also coherently added, has shown to be a good solution for increasing the resolution along the optical axis and for reducing the amplitude of the side lobes of the point spread function. However, the resolution in the transverse plane is poorer than in the case of one-photon excitation due to the larger wavelength involved in the two-photon fluorescence process. In this paper we show that a particular arrangement of the 4Pi microscope, referenced as 4Pi′ microscope, is a solution for obtaining a lateral resolution in the two-photon regime similar or even better to that obtained with 4Pi microscopes working in the one-photon excitation regime. 相似文献
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Chen I.-H. Chu S.-W. Sun C.-K. Cheng P.-C. Lin B.-L. 《Optical and Quantum Electronics》2002,34(12):1251-1266
Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. Thus induced two-photon absorption and the accompanied multi-photon absorption/ionization not only cause photo-bleaching but also cell damage in the vicinity of the focal point. In this paper, we study the wavelength dependent cell damage induced by high intensity femtosecond near infrared lasers. The study was performed with a Ti:sapphire laser and a Cr:forsterite laser. With a longer output wavelength from a Cr:forsterite laser, multi-photon absorption and auto-fluorescence were found to be significantly suppressed and the destructive plasma formation was found to be greatly reduced. Sustained multi-photon spectra can be observed in most plant specimens with a tightly focused Cr:forsterite laser beam under long term irradiation with more than 100 mW laser average power. In contrast, multi-photon absorption induced destructive plasma formation were frequently observed with a tightly focused Ti:sapphire laser beam within seconds with more than 10 mW laser average power. 相似文献
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荧光共焦扫描系统成像特性的优化 总被引:8,自引:0,他引:8
对荧光共焦扫描系统用光强点扩散函数进行傅里变换得到系统三维传递函数的数学模型,并由此求得环形透镜和各种有限大小探测器系统的光学传递函数。用计算机模拟和光学传递九数值计算,分析了采用不同环形透镜及探测器对系统成像特性的影响 相似文献
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在传统共聚焦显微技术的基础上,图像扫描显微技术使用面阵探测器来代替单点探测器,结合虚拟数字针孔并利用像素重定位和解卷积图像重构算法将传统宽场显微镜的分辨率提高一倍,实现了高信噪比的超分辨共焦成像.但是,由于采用逐点扫描的方式,三维成像速度相对较慢,限制了其在活体样品成像中的应用.为了进一步提高图像扫描显微术的成像速度,本文提出了一种基于双螺旋点扩散函数工程的多焦点图像扫描显微成像方法和系统.在照明光路中,利用高速数字微镜器件产生周期分布的聚焦点阵对样品进行并行激发和快速二维扫描;在探测光路中,利用双螺旋相位片将激发点荧光信号的强度分布转换为双螺旋的形式;最终,利用后期数字重聚焦处理,从单次样品扫描数据中重构出多个样品层的超分辨宽场图像.在此基础上,利用搭建的系统分别对纤维状肌动蛋白和海拉细胞线粒体进行成像实验,证明了该方法的超分辨能力和快速三维成像能力. 相似文献