用于双光子激发荧光寿命显微成像的高重复频率皮秒扫描相机

建立了一台基于新研制的高重复频率皮秒扫描相机的双光子激发荧光寿命显微成像系统,重点介绍所研制的高重复频率皮秒扫描相机。为了在高时间分辨力的同时扩大时间测量范围,实现大面积两维空间高时间分辨取样测量,从而提高采样速率和更有效地发挥扫描相机的作用,设计和研制了一种大面积、高时间分辨力扫描变像管和一种重复频率高达1MHz的斜坡电压扫描电路。基于上述关键部件所研制的扫描相机具有重复频率高、扫描速度可调、时间分辨力高、工作面积大、非线性低、触发晃动小等优点。用钛宝石飞秒激光器作为激光脉冲源,通过脉冲提取器将76MHz的高重复频率降低为1MHz,采用可调延时器和标准具对扫描相机的时间分辨力、扫描速度和非线性进行标定。该系统的时间分辨力达到6.5ps,非线性为2.60%,可测量的时间范围从十几皮秒到几十纳秒。测量了若丹明6G和香豆素314两种标准荧光染料的荧光寿命,取得了与参考文献一致的实验结果。     

时间维相位重建的改进算法

提出了一种时间维-相位重建的改进算法:利用折叠位相图进行时间相位展开以提高算法效率;推导并调整了基于时间相位展开算法初始条件从而减少所需的二维编码条纹图,增加了算法的实时性.从而可以更加有效地解决深度表面不连续和存在隔离物表面拓扑结构的景物相位重建问题.实验结果证明了本文所提出算法的可行性及有效性.     

圆扫描变像管人卫激光测距技术研究

在本文中,我们设计并研制了用于毫米级精度人卫激光测距的圆扫描时间分析系统。研制的圆扫描变像管理论时间分辨率可优于3ps,可用于双色测距;偏转灵敏度高达10cm/kV,偏转空间分辨率优于24lp/mm;利用双MCP内增强,增益～1×10⁶,并有门控功能以进一步提高信噪比和动态范围。扫描控制电路能提供300MHz、10W的正弦信号,其频率、幅度、相位稳定度均可满足要求。调试过程中较好地解决了阻抗变换、空间干扰和相位正交等关键问题。在仅2W的RF功率驱动下,即可实现稳定的φ24mm的圆扫描,系统实测时间分辨率达4.8ps。在实验室实测测距精度达亚毫米级。动态实验及模拟测距表明,该时间分析系统可满足第四代人卫激光测距系统的需要。     

Analysis of detection limit to time-resolved coherent anti-Stokes Raman scattering nanoscopy

In the implementation of CARS nanoscopy, signal strength decreases with focal volume size decreasing. A crucial problem that remains to be solved is whether the reduced signal generated in the suppressed focal volume can be detected. Here reported is a theoretical analysis of detection limit （DL） to time-resolved CARS （T-CARS） nanoscopy based on our proposed additional probe-beam-induced phonon depletion （APIPD） method for the low concentration samples. In order to acquire a detailed shot-noise limited signal-to-noise （SNR） and the involved parameters to evaluate DL, the T-CARS process is described with full quantum theory to estimate the extreme power density levels of the pump and Stokes beams determined by saturation behavior of coherent phonons, which are both actually on the order of ~ 109 W/cm2. When the pump and Stokes intensities reach such values and the total intensity of the excitation beams arrives at a maximum tolerable by most biological samples in a certain suppressed focal volume （40-nm suppressed focal scale in APIPD method）, the DL correspondingly varies with exposure time, for example, DL values are 103 and 102 when exposure times are 20 ms and 200 ms respectively.     

X射线光栅微分相衬成像视场分析

X射线光栅微分相衬成像对由轻元素构成的物质的内部探测具有传统吸收成像无法比拟的优势, 尤其在癌症的早期诊断和轻元素材料及器件的无损检测等领域应用潜力巨大. 大视场成像是影响该技术从实验室走向实际应用的重要因素. 针对大视场成像的客观需求, 基于菲涅耳衍射原理和光栅结构特征, 建立了量化物理模型用于分析影响成像视场的因素, 提出了实现大成像视场的有效途径, 为未来大视场光栅微分相衬成像方法的设计和应用提供理论依据.     

一种实现显微荧光寿命图的测量方法

将脉宽120fs、重复率76MHz激光引入激光扫描显微镜的激发光路,利用其扫描系统对荧光标记样品激发扫描,将激发出的荧光从荧光探测光路引入备用的外部探测口;在探测口接一快速光电倍增管,将光电倍增管的信号送给时间相关单光子计数器,获得时间相关的荧光强度图;最后通过计算机处理获得荧光寿命图。应用此系统对青色荧光蛋白(CFP)、黄色荧光蛋白(YFP)荧光寿命进行了测量,并应用CFP、YFP实现荧光共振能量转移的测量。通过实验看出利用已有的激光扫描显微镜,配合较先进的寿命测量方法,可以很好地实现显微荧光寿命图的测量。     

A new method of detecting interferogram in differential phase-contrast imaging system based on special structured x-ray scintillator screen

An x-ray scintillator screen with a special structure,functioning as detector and analyser grating,was proposed for collecting the interferogram of differential phase contrast imaging without absorption grating and difficulty of fabrication by a state of the art technique.On the basis of phase grating diffraction,a detecting model of the scintillator screen was built for analysing the phase and absorption information of objects.According to the analysis,a new method of phase retrievals based on two-images and the optimal structure of screen were presented.     

硬X射线相位光栅的设计与研制

针对在普通实验室和医院实现40—100keVX射线相衬成像的需求,考虑到成像系统参数、X射线源空间相干特性及光栅衍射效率,设计出硅基相位光栅结构参数.利用我们已发展的光助电化学刻蚀技术研制出直径为5英寸的相位光栅,其空间周期为5.6μm,线宽为2.8μm,深度为40—70μm.在理论分析的基础上,通过提高硅片两端有效工作电压和修正Lehmann电流密度公式,解决了实际刻蚀过程中出现的钻蚀问题.由实验结果可知,本方案对制作大面积高精度相位光栅十分有效。     

Improving the precision of fluorescence lifetime measurement using a streak camera

Streak camera has high temporal resolution and high sensitivity, and is a powerful tool in biomedical study to measure fluorescence lifetime and perform fluorescence lifetime imaging. However, nonuniformity of the gain in the streak tube and nonlinearity of the sweeping speed limit the precision of fluorescence lifetime measurement, particularly when fluorescence lifetimes are short. We have constructed a two-photon excitation fluorescence lifetime measurement system that is based on a synchroscan streak camera and have developed accordingly a method to correct the effect of gain nonuniformity and nonlinearity of sweeping speed on the measurement precision. A continuous-wave laser of high stability is used to calibrate the gain of the streak camera, and a Fabry-Perot etalon is used to calibrate the nonlinearity of the sweeping speed. Fitting algorithms are used to correct the gain of the streak camera and nonlinearity of the sweeping speed respectively, which significantly improves the measurement precision of the system, as characterized through the fluorescence lifetime of the short-lived fluorescence dye, Rose Bengal. Experimental results show that the measurement fluctuation of the lifetime has been improved from more than 10% to 2% after correcting the effects of gain nonuniformity and sweeping speed nonlinearity.     

圆锥形花样偏转器

国内目前广泛使用的静电偏转系统是平行或斜行金属板结构。为了实现两维偏转,必须采用两对相继正交的偏转板。它的缺点是很明显的:1)由于第二对偏转板不可能置于交叉点处,偏转板之间的距离就要加大,从而导致偏转灵敏度降底,偏转象质也会变坏;2)占据了更大的等位空间,装架带来更大的同轴难度。