激光诱导时间分辨固体表面荧光光谱系统

采用斜射式激发样品光路结构,设计了应用于测试时间分辨固体表面荧光光谱系统。利用该系统研究了氧化锌薄膜在355 nm激光脉冲激发下的时间分辨固体表面荧光光谱。结果表明,该系统能够实现测量时间分辨固体表面荧光光谱,并由此测量了样品的固体表面荧光寿命。该系统结构简单可靠,灵敏度高,响应速度快。     

基于激光诱导叶绿素荧光寿命成像技术的植物荧光特性研究

针对植物荧光遥感探测中信号易受干扰的问题, 提出了一种用于评估植物生长状况及环境监测的荧光寿命成像技术. 采用凹透镜对355 nm波长的激光扩束, 再照射植物激发叶绿素荧光, 由增强型电荷耦合器件接收荧光信号. 采用时间分辨测量法, 连续用相同激光脉冲照射植物以激发相同的荧光信号, 同时不断改变激光脉冲触发探测器启动的延时时间, 从而能够得到完整的离散荧光信号分布图像. 对植物特定位置点产生的离散荧光信号进行拟合, 再运用一种改进型的迭代解卷积法可反演高精度的荧光寿命; 进而反演图像各点的荧光寿命以生成植物的荧光寿命分布图. 该方法所绘制的荧光寿命图比荧光强度图能更准确地反映植物内部的叶绿素含量, 并对活体植物叶绿素荧光寿命的物理特性进行了初步研究, 证明叶绿素荧光寿命与植物生理状态存在一定关联; 并且叶绿素荧光寿命与活体植物所处环境存在着复杂的关系. 未来将与生物物理学家们合作, 继续探寻叶绿素荧光寿命与植物生存环境的关系.     

双光子阵列点激发同时多维荧光信息的处理

五维同时荧光信息显微成像方法是一种新的荧光信息获取技术，它采用了双光子阵列点激发方式.这一方法可同时获取激发阵列点每点荧光的位置信息、荧光光谱信息和荧光寿命信息，弥补了现有荧光检测技术的不同功能信息不具有同时性的缺陷.给出了从这种技术的复合信息中提取复合光谱几何强度结构图像、不同光谱几何强度结构图像、不同光谱寿命图像的方法.提出了一种激发荧光强度修正系数矩阵方法，消除阵列点激发光强不均匀对激发荧光强弱产生的不利影响，取得明显效果.实验对实际样品做了数据采集和处理，给出图像结果，表明处理的效果良好.对存在的问题也作了讨论. 关键词： 荧光信息处理 双光子 荧光光谱 荧光寿命     

基于CIE色度图的博尔塔拉河与精河水体荧光峰发光性初探

以艾比湖主要入湖河流博尔塔拉河及精河水体为研究对象,使用寻峰法找出水体荧光峰,采用色坐标分析法对博尔塔拉河与精河水体荧光发射光谱特性及荧光峰的发光性进行分析。首先,博尔塔拉河与精河水体的荧光发射光谱均含有3个荧光峰,各荧光峰的出现位置与峰强度大小均不同。博尔塔拉河第二荧光峰远大于第一荧光峰或第一荧光峰与第二荧光峰基本持平;精河前5个样点的3个荧光峰强度随波长增大呈依次递减分布,6号采样点第一荧光峰与第二荧光峰基本持平。其次,博尔塔拉河与精河3个荧光峰在色坐标中分布位置大致相同,且各点的荧光峰分布较为聚集,均在蓝光区域,属蓝光发射。最后,各荧光峰在CIE坐标中聚集分布,第一荧光峰在色坐标最底端;第二荧光峰分布在第一荧光峰上端,x坐标与第一荧光峰接近;第三荧光峰整体聚集分布在蓝光区域的右上角。     

Improved Routine Bio-Medical and Bio-Analytical Online Fluorescence Measurements Using Fluorescence Lifetime Resolution

Fluorescence techniques are widely used as sensitive detection methods in bio-analytics. The use of the bio-physical parameter fluorescence lifetime additional to the spectral characteristics of fluorescence has the potential to improve fluorescence-related detection methods in terms of selectivity in signal recognition, robustness against disturbing influences, and the accessibility of novel bio-chemical process parameters. This article describes the technical set up of a time-resolving instrument with either a fixed time-gated detection principle for improved evaluation of tissue metabolism by an online monitoring of the tissue autofluorescence or a direct fluorescence lifetime detection principle for lifetime-based fluorescent assays.     

氧化镍位于510～610 nm的激光诱导荧光光谱 (cited: 1)

采用激光诱导荧光技术在510～650 nm研究了超声射流冷却下的氧化镍光谱,观测了55个振转谱带. 通过转动分析发现激发态的同位素位移、振转间隔和转动常数都表现出高度的不规则性,26个[Ω=0,1]-X³∑^-₀的跃迁谱带被暂时归属为5个振动序列. 此外实验中还观测了部分谱带的色散谱和寿命.     

Polarization of Fluorescence of Excimer Vinylpyrene

The polarization of the excimer luminescence of copolymers of 1-vinylpyrene with methyl methacrylate has been studied. The obtained polarization dependences in absorption and fluorescence spectra differ for luminescence in the region of the maximum of the excimer band and of its longwave portion, which is due to the influence of the nonequilibrium configurations of the dimeric complex on the polarization properties of its luminescence.     

海洋微藻生长过程藻液三维荧光特征

利用三维荧光光谱技术监测了中肋骨条藻（Skeletonema costatum）,塔玛亚历山大藻（Alexandrium tamarense）,微小亚历山大藻（Alexandrium mimntum）,锥状斯氏藻（Scrippsiella trochoidea）,东海原甲藻（Prorocentrum donghaiense）及海洋原甲藻（Prorocentrum micans）实验室培养过程中产生的荧光溶解有机物。中肋骨条藻为代表的硅藻主要产生类腐殖质荧光物质,而甲藻在指数增长期主要产生类蛋白荧光物质。进入消亡期后类蛋白荧光和类腐殖质荧光强度迅速增大,原因可能是衰老、死亡藻细胞的破碎释放出大量的荧光有机物质所致,此外还有细菌对非荧光有机物进一步降解。塔玛亚历山大藻、微小亚历山大藻、东海原甲藻及海洋原甲藻的类蛋白荧光强度在消亡后期由于细菌降解或光降解等因素而降低。同属微藻产生的荧光物质相似,例如塔玛亚历山大藻与微小亚历山藻、东海原甲藻与海洋原甲藻,但具体荧光峰位置有所不同。     

光参量荧光寿命分布

 根据荧光寿命定义和放大过程获得的增益方程,研究了新型非线性晶体BaAlBO₃F₂中三种不同的相位匹配模式下中心波长532 nm泵浦的参量荧光光子寿命的分布特性。结果表明,当注入单色信号光时,参量荧光寿命随泵浦光相位匹配角的增加由椭圆体分布变化为圆环体分布。采用宽带信号光时,则由分散分布改变为集中在近200 nm波长范围内分布,并随泵浦光相位匹配角的增加而逐步减小。考虑宽带泵浦光注入情况时,参量荧光寿命的分布范围由于相位匹配范围增加而随之扩展。     

Studies on Molecular Structure of Ethanol-Water Clusters by Fluorescence Spectroscopy

It was found that ethanol and water molecules mixed together can form new clusters. Based on the steady state spectral characteristic and the mole ratio method, three possible bonding constants of ethanol-water clusters were deduced. To confirm the structure of these clusters, the fluorescence lifetimes of different emission bands were investigated, and the average lifetime of the entire decay process was found to vary as the volume percent of ethanol changed. Furthermore, the possible bonding constant n of molecular clusters was determined to be 2 based on the time-resolved spectra, and the cluster was concluded to be a chain structure formed by one ethanol molecule and two water molecules with hydrogen bonds.     

Steady-State and Time-Resolved Fluorescence Polarization Behavior of Acenaphthene

Steady-state and time-resolved fluorescence polarization studies have been carried out on acenaphthene (ACE) in low-temperature glass solutions and at room temperature. In the low-temperature glass the fluorescence polarization values vary considerably with both emission and excitation wavelength. There is a time dependence (on the nanosecond time scale) of the fluorescence anisotropy, r(t), at 77 K, which has a strong dependence upon the excitation and emission wavelengths. Under these conditions, the time-dependent decay of the anisotropy is not attributable to chromophoric motion. The observations are consistent with emission from two closely lying and interconverting excited states. Rate constants for the photophysical processes involved have been determined by fitting the data using a model proposed by Fleming et. al. The results are discussed with particular reference to the care required in using dynamic fluorescence polarization measurements to determine energy transfer rates in systems containing this chromophore.     

脉冲激光激发的原子共振荧光

本文从含有弛豫项的光与二能级原子相互作用的布洛赫(Block)方程出发,利用计算机进行数值求解,在强共振激发条件下,给出了几种不同的激光脉冲(包括常见的高斯脉冲)激发的原子共振荧光谱,获得了原子在瞬态辐射过程中的一些新现象。     

Spectral characteristics of luminescence of solutions of complex molecules under conditions of electronic excitation transfer to a saturable acceptor in the case of inhomogeneous broadening of electronic energy levels

Spectral characteristics of solutions of complex molecules under conditions of inhomogeneous broadening of energy levels are considered in the case when the nonlinear dependence of the population of molecular states on the excitation intensity is determined not by saturation of molecular levels but by exchange of the electronic excitation energy with the environment. Calculations have shown that the dependence of the position of the fluorescence spectrum on the excitation frequency is nonmonotonic in solutions of the type and varies substantially with the excitation intensity. Belarusian State University, 4, F. Skorina Ave., Minsk, 220072, Belarus. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 64, No. 2, pp. 164–168, March–April, 1997.     

用时间分辨光谱研究红细胞悬液的荧光发射

实验获得了激光照射红细胞悬液的荧光光谱,并分别监测不同荧光峰值波长处强度随时间的衰变过程,测试了其相应的荧光寿命。结果表明,在波长为407nm的激光照射下,红细胞悬液向外发射中心波长分别位于596,628,692nm的荧光光谱,各荧光峰对应衰变过程的平均荧光寿命分别为1.97,13.31,14.58ns。利用荧光强度和吸收率的加和性表示了混合物的总吸收率和总荧光发射强度,通过理论计算获得了红细胞悬液中锌卟啉、原卟啉和其他游离物参与荧光发射的相对含量和相对强度在不同荧光峰位的变化关系,进一步解释了不同峰位处荧光发射强度和平均荧光寿命的变化原因。     

Voltage-Gated Metal-Enhanced Fluorescence

We demonstrate the influence of electrical current on the ability of surface plasmons to amplify fluorescence signatures. An applied direct current across Silver Island Films (SIFs) of low electrical resistance perturbs the fluorescence enhancement. For a given applied current, surface plasmons in just-continuous films are sparsely available for fluorophore dipole-coupling and hence the enhanced fluorescence is gated as a function of the applied current. For thicker, low resistance films, sufficient charge carriers are now present in the metal that metal-enhanced fluorescence (MEF) is perturbed to a lesser extent, induced surface plasmons readily formed on the surface by the close-proximity dipole.     

Video fluorescence microscopic techniques to monitor local lipid and phospholipid molecular order and organization in cell membranes during hypoxic injury

Digitized video microscopy is rapidly finding uses in a number of fields of biological investigation because it allows quantitative assessment of physiological functions in intact cells under a variety of conditions. In this review paper, we focus on the rationale for the development and use of quantitative digitized video fluorescence microscopic techniques to monitor the molecular order and organization of lipids and phospholipids in the plasma membrane of single living cells. These include (1) fluorescence polarization imaging microscopy, used to measure plasma membrane lipid order, (2) fluorescence resonance energy transfer (FRET) imaging microscopy, used to detect and monitor phospholipid domain formation, and (3) fluorescence quenching imaging microscopy, used to spatially map fluid and rigid lipid domains. We review both the theoretical as well as practical use of these different techniques and their limits and potential for future developments, and provide as an illustrative example their application in studies of plasma membrane lipid order and topography during hypoxic injury in rat hepatocytes. Each of these methods provides complementary information; in the case of hypoxic injury, they all indicated that hypoxic injury leads to a spatially and temporally heterogeneous alteration in lipid order, topography, and fluidity of the plasma membrane. Hypoxic injury induces the formation of both fluid and rigid lipid domains; the formation of these domains is responsible for loss of the plasma membrane permeability barrier and the onset of irreversible injury (cell death). By defining the mechanisms which lead to alterations in lipid and phospholipid order and organization in the plasma membrane of hypoxic cells, potential sites of intervention to delay, prevent, or rescue cells from hypoxic injury have been identified. Finally, we briefly discuss fluorescence lifetime imaging microscopy (FLIM) and its potential application for studies monitoring local lipid and phospholipid molecular order and organization in cell membranes.     

Lanthanide Complex-Based Fluorescence Label for Time-Resolved Fluorescence Bioassay

Different from organic fluorescence dyes, fluorescent lanthanide complexes have the fluorescence properties of long fluorescence lifetime, large Stokes shift and sharp emission profile, which makes them favorable be used as the fluorescent labeling reagents for microsecond time-resolved fluorescence bioassay. Lanthanide complex-based fluorescence labels have been successfully used for highly sensitive time-resolved fluorescence immunoassay, DNA hybridization assay, cell activity assay, and bioimaging microscopy assay. Since the technique allows easy distinction of the specific fluorescence signal of the long-lived label from short-lived background noises associated with biological samples, scattering lights (Tyndall, Rayleigh and Raman scatterings) and the optical components (cuvettes, filters and lenses), the sensitivity of fluorescence bioassay has been remarkably improved. This paper summarized the recent developments of lanthanide complex-based fluorescence labels and their applications in time-resolved fluorescence bioassays mainly based on the authors’ researches and relative publications.     

激光诱导荧光寿命及其测量

激光诱导荧光特性的研究可用于包括心血管病在内的多种疾病的诊断。荧光发射包括光谱（频域）和时间（时域）两方面的信息，后者表现为荧光寿命。在很多情况下，测量荧光寿命是比测量光谱更为有效的诊断方法。本文从理论上讨论了荧光寿命问题，并介绍两种测量方法，可用于测量人体正常组织和病变组织的激光诱导荧光寿命     

Influence of a Solvent on the Quantum Yield and Duration of the Fluorescence of Tetraazoporphin

In view of the discrepancies between the values available in the literature for the photophysical parameters of tetraazaporphin in solutions, we measured its quantum yield and duration of fluorescence in a number of solvents. It has been found that alcohols (isobutanol and isopropanol) quench the fluorescence appreciably. For a solution in toluene — a stable chemically inactive and low-polar solvent — the measured quantum yield of the fluorescence of tetraazaporphin is equal to 0.180 ± 0.015 and its duration is 3.4 ± 0.1 nsec. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 5, pp. 697–699, September–October, 2005.     

双酰胺稀土配合物的合成及荧光性能

为寻求发光性能良好的配合物,设计合成了一种新的双酰胺配体1,4-二 苯(L),并在氯仿和乙酸乙酯溶液中合成了其与硝酸钐、硝酸铕和硝酸铽的发光稀土配合物。元素分析数据表明稀土硝酸盐与配体形成的是 1 : 1 型的配合物;红外光谱显示配合物具有相似的配位结构。对配体及其配合物的荧光进行了详细的研究,结果表明:钐、铕和铽配合物分别呈现出Sm³⁺、Eu³⁺、Tb³⁺的特征发射,铕离子处于不对称中心格位,硝酸铽配合物荧光相对强度最大。