Evaluation of the inhibitory effect of docosahexaenoic acid and arachidonic acid on the initial stage of amyloid β1-42 polymerization by fluorescence correlation spectroscopy

Amyloid β(Aβ)1-42 fibrillation is a crucial step in the development of pathological hallmarks, such as neuritic plaques and neurofibrillary tangles, of Alzheimer’s disease (AD). In this study, we evaluated the effects of free docosahexaenoic acid (DHA), an essential brain polyunsaturated fatty acid (PUFA), on the inhibition of Aβ1-42 fibrillation by fluorescence correlation spectroscopy (FCS), a technique capable of detecting molecular movements and interactions in solution. We also examined whether free arachidonic acid (AA), eicosapentaenoic acid (EPA), and metabolites of DHA, including neuroprotectin D1 (NPD1, 10S, 17S-dihydroxy-DHA), resolvin D1 (RvD1, 7S, 8R, 17S-trihydroxy-DHA), and didocosahexaenoyl glycerol (diDHA), affect Aβ1-42 polymerization. The results of the FCS study reveal that DHA and AA significantly reduced the diffusion time of TAMRA (5-carboxytetramethylrhoda-mine)-Aβ1-42 by 28% and 31%, respectively, while EPA, NPD1, RvD1, and diDHA had no effects on diffusion time. These results indicate that DHA and AA inhibited Aβ1-42 polymerization and that their inhibitory effects occurred at the initial stage of Aβ1-42 polymerization. This study will advance the research on PUFAs in preventing AD progression.

Combination of Micro-fluidic Chip with Fluorescence Correlation Spectroscopy for Single Molecule Detection

A single molecule detection technique was developed by the combination of a single channel poly （dimethylsiloxane）/glass micro-fluidic chip and fluorescence correlation spectroscopy （FCS）. This method was successfully used to determine the proportion of two model components in the mixture containing fluorescein and the rhodamine-green succinimidyl ester.

Observation of Cell-Size Variation under Environmental Stress by Fluorescence Correlation Spectroscopy without Objective Image Magnification

Fluorescence correlation spectroscopy （FCS） without objective image magnification （without using con-focal microscope） was applied to observe the variation in cell size of Escherichia coli （E. coli） induced by the anti-cancer agent MitomycinC （MMC）. In the system without image magnification followed in this study, the suspension of E. coli cells was stirred, and the difference in movement due to the different cell sizes induced by the compulsive solution flow was detected. The addition of 0.1-0.4 pg/L of MMC elongated the E. coli cell length from about 3.6 to 7.8μm. The flow cell （i.d. = about 1 mm） also produced a size-dependent correlation curve, The present system is not based on single molecular FCS but is inexpensive and effective at observing the variation in cell size induced by environmental changes.

荧光相关光谱(FCS)在生物活细胞中的应用

除了传统的分子浓度和扩散时间的测量荧光相关光谱(FCS),还可测量分子动力学或化学反应的动力参数.但是由于分子相互作用网络的复杂性,FCS在生物活细胞中的应用还有许多问题和困难需要解决.探讨了FCS在生物活体中分子相互作用网络、细胞膜、膜表面受体、细胞内结构分析以及胞内动力学研究等方面的应用.

Fluorescence correlation spectroscopy in laser gradient field

Fluorescence correlation spectroscopy (FCS) is capable of probing dynamic processes in living biological systems. From photon fluctuation of fluorescing particles which diffuse through a small detection volume, FCS reveals information on the concentration and the structure of the particles, as well as information on microscopic environment. In this note, we study the radiation forces experienced by Rayleigh particles in a laser field in details, and analyze the effects of gradient field on FCS measurements.

Quantification of Membrane Protein Dynamics and Interactions in Plant Cells by Fluorescence Correlation Spectroscopy

Deciphering the dynamics of protein and lipid molecules on appropriate spatial and temporal scales may shed light on protein function and membrane organization. However, traditional bulk approaches cannot unambiguously quantify the extremely diverse mobility and interactions of proteins in living cells. Fluores- cence correlation spectroscopy （FCS） is a powerful technique to describe events that occur at the singlemolecule level and on the nanosecond to second timescales; therefore, FCS can provide data on the heterogeneous organization of membrane systems. FCS can also be combined with other microscopy techniques, such as super-resolution techniques. More importantly, FCS is minimally invasive, which makes it an ideal approach to detect the heterogeneous distribution and dynamics of key proteins during development. In this review, we give a brief introduction about the development of FCS and summarize the significant contributions of FCS in understanding the organization of plant cell membranes and the dy- namics and interactions of membrane proteins .We also discuss the potential applications of this technique in plant biology.

Quantitative deconvolution of autocorrelations and cross correlations from two-dimensional lifetime decay maps in fluorescence lifetime correlation spectroscopy

Fluorescence correlation spectroscopy （FCS） is a widely used method for measuring molecular diffusion and chemical kinetics. However, when a mixture of fluorescent species is taken into account, the conven- tional FCS method has limitations in extracting autocorrelations for different species and cross correla- tions between different species. Recently developed fluorescence lifetime correlation spectroscopy （FLCS） based on time-tagged time-resolved （TITR） photon recording, which can record the global and micro arrival time for each individual photon, has been used to discriminate different species according to fluorescence lifetime. Here, based on two-dimensional lifetime decay maps constructed from TITR photon stream, we have developed a quantitative lifetime-deconvolution FCS model （LDFCS） to extract precise chemical rates for chemical conversions in multi-species systems. The key point of LDFCS model is separation of different species according to the global distribution of fluorescence lifetime and then deconvolution of autocorrelations and cross-correlations from the two-dimensional lifetime decay maps constructed bv the micro arrival times of photon pairs at each delay time.

Advantage of Fluorescence Correlation Spectroscopy for the Study of Polyelectrolytes

The advantage of fluorescence correlation spectroscopy to study single chain behavior of polyelectrolytes has been demonstrated by checking the coil-to-globule transition ofpoly 2-vinylpyridine with the change ofpH value in aqueous solution. The ultra-high sensitivity of FCS allows measurement at extreme dilution where the effect of electrostatic interaction between the chains is greatly suppressed. The results exposed first-order conformation tran- sition of P2VP as detected by FCS while inter-chain aggregation occurred in the experiments of dynamic light scat- tering.

基于光谱特征的多重金属离子与DOM结合特性

通过淬灭滴定实验,文章采用同步荧光光谱和傅里叶红外光谱(FTIR)结合二维相关光谱分析,探究Cu^(2+)和Cd^(2+)多重离子与溶解态有机质(DOM)结合特性。研究结果表明,(1)DOM与Cu^(2+)和Cd^(2+)结合稳定常数(lg KM)分别为4.75~5.18和3.71~4.12,Cu^(2+)与DOM的结合能力要强于Cd^(2+)。(2)Cd^(2+)浓度为0和100μmol/L时,DOM点位与Cu^(2+)的结合顺序均为430~500 nm>330~420 nm,且Cd^(2+)存在时Cu^(2+)与DOM的lg KM为4.56~4.95。对比结果表明Cd^(2+)的存在并未影响DOM点位与Cu^(2+)的结合顺序,但减弱了DOM与Cu^(2+)的结合能力。(3)FTIR结果表明,Cd^(2+)浓度为0和100μmol/L时DOM官能团与Cu^(2+)结合顺序依次为酚类C-O>酰胺N-H>酰胺C=O>羧基C=O>芳香族C-H>多糖C-O和酰胺N-H>酰胺C=O>酚类C-O>芳香族C-H>多糖C-O。Cd^(2+)与酚类C-O结合后影响了Cu^(2+)与DOM中酚类C-O和羧基C=O官能团的结合。水环境中多重金属离子共存时的生物有效性和生态风险应受到关注。

Fluorescence fluctuation spectroscopy and its artifacts:simulations and tests

Fluorescence fluctuation spectroscopy (FFS) technique is capable of monitoring changes in concentration, mass, size and structure of fluorescent-labeled bio-molecules in microscopic volume and is suitable for measuring biological interactions in living cells. FFS data may be affected by many experimental factors in complicated biological systems. Using a Monte Carlo approach, we generate fluorescence fluctuation data for different experimental systems. This approach helps to separate the contributions by different experimental factors in a complicated fluorescence fluctuation spectrum. It also helps to validate new theoretical models and new fitting formulations. We describe the algorithm of the simulation program and tests on its statistical performance. The program is then used successfully to study the effects of several experimental factors on FFS detection.

不同有机肥对生菜生育期土壤剖面DOM分布的影响

施用有机肥是农作物生产过程中提质增效的有效途径,研究不同来源有机肥施加下土壤剖面溶解性有机质(DOM)的分布规律有助于优化有机肥利用及进一步了解DOM环境行为。采用二维相关光谱(2DCOS)等方法,研究施加不同来源有机肥(猪粪、鸡粪、羊粪、牛粪和沼渣)对土壤剖面DOM分布规律的影响。结果显示:有机肥还田主要影响土壤DOM的相对含量及组分构成。施加不同来源有机肥后,土壤中DOM相对含量始终大于对照(CK)处理,其中0~10cm土层DOM相对含量影响最大,平均增加14.67g·kg-1。不同类型有机肥对增加剖面DOM有一定差异,其中鸡粪有机肥对剖面DOM增加影响最大,增加了21.42%。有机肥的施加主要提高了类酪氨酸组分(C4)的相对含量,对10~20cm土层C4相对含量的影响最大,平均增加4.12%,其中羊粪和沼渣有机肥影响最大,分别增加了7.80%、7.89%。2D-COS分析显示,不同来源有机肥施用对溶解性微生物代谢产物、类蛋白质组分(295nm、315nm)影响最大,CK、鸡粪有机肥处理主要影响的是溶解性微生物代谢产物组分,其他处理最先响应的是类蛋白质组分;不同来源有机肥施用下,腐殖化指数HIX和Fn(330)、Fn(280)均随土层深度的增加而减小,但HIX和Fn(330)、Fn(280)的变化随有机肥种类不同而有所差异,其中牛粪有机肥对HIX和Fn(330)影响最大,羊粪有机肥对Fn(280)影响最大。生物指数BIX和新鲜度指数β∶α随着土层深度增加而增加,其中沼渣的影响最显著,但荧光指数FI并无显著变化。

荧光共振能量转移技术在生命科学中的应用及研究进展

荧光共振能量转移(FRET)是指两个荧光基团间能量通过偶极-偶极耦合作用以非辐射方式从供体传递给受体的现象,可被用于测定分子间的距离。FRET荧光探针主要有荧光蛋白、有机荧光染料、镧系染料和量子点,近年来有许多新的应用;测试方法上与时间分辨、单分子检测、荧光寿命和荧光相关光谱等技术联用取得了更好的成效。

土壤粒径大小对蒽荧光特性的影响及校正

为了准确检测土壤中的多环芳烃,以土壤中典型多环芳烃污染物蒽为检测对象,研究了土壤粒径大小对其荧光特性的影响,并提出了一种校正土壤粒径大小对多环芳烃标准曲线影响的方法。研究了蒽在土壤中的荧光特性,指出蒽在421nm、442nm和470nm处出现较强的荧光峰。接着,制备7种不同粒径大小的蒽土壤样品,并以土壤粒径大小为外扰,构建了同步和异步二维相关荧光谱,研究了蒽荧光强度和304nm处瑞利散射光强随土壤粒径大小的变化。结果显示,随着土壤粒径增大,蒽荧光强度和304nm处瑞利散射光强度都有增强。最后,分别建立了80目和160目土壤粒径下定量分析土壤蒽浓度的标准曲线,并通过304nm处瑞利散射光对蒽荧光进行校正。结果表明:该方法有效降低了土壤粒径大小对蒽标准曲线的影响。

二维相关荧光光谱鉴别4种食用植物油种类的研究

采用Cary Eclipse荧光分光光度计检测了4种食用植物油的同步荧光光谱,结合加热时间扰动下的二维相关荧光光谱技术,对一级大豆油、花生调和油、芝麻油和棕榈油,进行了快速无损的识别方法的研究。结果表明,在同步荧光光谱图上,不同植物油的特征吸收峰在出峰数目、出峰位置和峰强上均存在一定的差异;而在二维相关荧光谱图上,由于4种植物油的自动峰及交叉峰的位置、数量和强度不同,其差别体现得更为明显和直观。因此,二维相关图谱法可以作为鉴别不同植物油种类的一种有效方法。

同步荧光光谱技术研究胶原基表面活性剂溶液中分子的聚集行为

基于胶原基表面活性剂（collagen-based surfactant,CBS）中酪氨酸（Tyr）和苯丙氨酸（Phe）的荧光特性,应用恒波长差（Δλ）为15nm的同步荧光光谱技术研究CBS浓度、溶液pH值、NaCl浓度和温度对其在水溶液中分子的聚集行为的影响,并以温度为外扰,利用二维同步荧光相关分析研究CBS分子中Tyr残基和Phe残基随温度变化的响应顺序。结果表明,CBS分子在261和282nm处出现了分别归属于Phe和Tyr的特征吸收峰。随着CBS浓度的升高,CBS分子中Phe残基和Tyr残基数量逐渐增多使CBS分子聚集程度增加,并导致荧光强度增强;CBS溶液pH值（pH 5.0）在等电点附近时,由于CBS分子的疏水作用和氢键形成能力加强,导致CBS分子聚集程度增强;CBS溶液中NaCl浓度的升高,则使CBS分子间排斥力减弱,从而导致CBS分子的聚集;然而温度升高,CBS由聚集状态逐渐变为单分子状态,因猝灭、变性以及氢键形成能力降低,荧光强度逐渐降低。以温度为外扰的二维同步荧光相关光谱分析可知：低温下（10^40℃）,CBS聚集体随温度升高逐渐松散,位于聚集体内部的Phe残基比Tyr残基优先响应;而在45~70℃时,CBS由单分子状态逐步水解为无规卷曲构造,Tyr残基的间距变大,氢键形成能力大大降低,Tyr残基比Phe残基优先响应。

荧光相关光谱及其在单分子检测中的应用进展

单分子检测在生命科学、化学、物理学等领域具有重要的意义。荧光相关光谱是单分子检测的新技术,在生命科学领域有巨大的应用潜力。综述了荧光相关光谱单分子检测的原理、实验技术以及在生物分子相互作用、活细胞、核酸、疾病诊断、高通量筛选以及与毛细管电泳联用等领域的研究,并展望了其发展前景。

通过DOM的光谱特性对长江源头典型高原深水湖泊进行评价

天然溶解有机质（dissolved organic matter,DOM）研究在水体生态环境系统的修复研究中扮演着重要的角色,是近年来水环境研究领域的一个热点和难点。目前,紫外？可见光谱和分子荧光光谱在天然溶解有机质是表征天然溶解有机质的重要有效的手段,也是研究天然溶解有机质的主要手段。红枫湖为贵阳市5个主要饮用水源地之一,对贵阳市社会发展具有重要作用。采用紫外？可见光谱和分子荧光光谱相结合的研究方法,对采自红枫湖中10个点的水样中天然溶解有机质的光谱特性研究。研究结果表明,红枫湖水体光谱特征主要表现为紫外？可见吸光值参数E3/E4的变化范围是1.70~8.77。除后五火电厂外,水下10 m比水下5 m的E3/E4值要高。此外,荧光指数f450/500的变化范围是1.38~1.52。初步认为红枫湖目前的主要有机物污染是陆源性的。后五火电厂,北湖湖心和张官三点水样的光谱特征与其他地方不同。同时还表明溶解有机质的腐殖化程度与水深具有一定的相关性。

基于荧光光谱的茶汤中拟除虫菊酯类农药的智能识别

基于三维荧光光谱和二维荧光相关光谱,对茶汤中两种拟除虫菊酯农药进行了识别。利用图像局部极值算法,分别提取出三维荧光光谱图和二维相关光谱图中的峰、谷位置信息,然后进行峰位匹配实现茶汤中农药组分的智能识别。结果表明,三维荧光光谱图受到荧光峰重叠的影响,不能对氰戊菊酯进行有效匹配。而利用二维相关光谱不仅可以克服这一缺点,而且不受混合物中农药组分浓度的影响。基于二维荧光相关光谱,对实际茶汤中氰戊菊酯和顺式氯氰菊酯进行了识别,识别率分别为80%和83%。

二维相关荧光光谱技术

从发展历史、计算方程、一般规则和特有性质等方面系统地介绍了近年来在二维相关荧光光谱技术方面的方法探索和应用进展。以不同的外扰方式,如浓度、激发波长、猝灭以及其他外扰方式如pH等分类,举例阐述了二维荧光相关光谱的可操作性及其应用,并与普通一维荧光光谱比较,说明了二维荧光相关光谱技术的优势。

菜籽油热氧化过程中荧光光谱特征的变化研究

为探讨菜籽油热氧化过程中荧光光谱特征的变化规律,采用平行因子分析方法结合二维相关技术解析菜籽油的三维同步荧光光谱,并与油脂氧化化学指标进行相关分析。结果表明:菜籽油具有叶绿素a的荧光特征(Δλ=10 nm,λex=674 nm)和叶绿素b的荧光特征(Δλ=60 nm,λex=620 nm);热氧化过程中,产生了(Δλ=60 nm,λex=448 nm)的特征同步荧光峰。双因子的平行因子分析表明:Δλ为10和60 nm的同步荧光光谱分别为第一、第二主要成分,分别对应叶绿素a和油脂氧化产物的荧光特征;随着热氧化时间的延长,叶绿素a的同步荧光强度降低,而油脂氧化产物的同步荧光强度逐渐升高。二维相关分析表明,叶绿素a热氧化速度、叶绿素b热氧化速度、油脂氧化产物生成速度依次降低。代表叶绿素氧化程度的SFI674和代表油脂氧化产物产量的SFI448与油脂氧化化学指标具有良好的相关性。因此,同步荧光光谱可作为衡量食用油热氧化劣变的指标。