鸡胚完整脑的光子发射和光激发光研究

利用自制的单光子记录系统,对离体的浸于DMEM培养液中的鸡胚的完整脑所进行的光子发射测量显示出整脑与培养液相比有光子发射.完整脑的光子发射强度与胚龄,鸡胚的健康状况和离体后脑组织的新鲜程度有关.在对完整脑的光激发光测量中观察到一种短寿命的延迟发光,它的衰减过程不服从指数衰减规律而表现出双曲线衰减行为.从生物光子发射归因于生物体内的非定位的相干性电磁场及Frohlish的生命系统中存在着长距离的相干性的理论,讨论了利用生物光子发射研究细胞间通讯、生物的调节及生物与环境关系的意义,以及所检测的光子发射与生物系统自身的相干性电磁场的关系,并提出应从环境对生物系统自身场的影响来理解所测的生物光子发射.     

淡水鱼超微弱光子辐射的实验研究

采用一台高灵敏的生物活体单光计数系统对淡水鱼的主要超微弱发光特性进行了研究.淡水鱼的各种不同脏器的光子辐射强度数值为3.6—97.3CPS.发射光谱的最大值接近640nm处和460nm附近.实验结果表明,超微弱光子辐射的总强度随所测的时间而变化,加入UOUO_2~( )离子之后,其发光强度下降.     

不同光波长下大光金鱼花叶片超微弱发光的光子计数统计

用生物超微弱发光探测技术,对植物叶片的超微弱发光透过滤波片进行测量,并用量子光学中的光子计数分布统计方法得到的结果表明,不同波长下,大光金鱼花叶片的光子计数分布不同,但幼叶和老叶的光子计数分布的规律基本相同,而成熟叶片光子计数分布的规律比较紊乱。     

植物幼苗原发性超弱光子成像的研究

采用光子计数成像系统（PIAS）对植物幼苗萌发过程的超弱发光进行观察。结果表明,自发光子长时间积累可形成二维图象;光子计数和采集图象均可得到植物体的自发发光;通过实验探测到幼苗的根,叶在同一平面图象有不同的发光表现;光子成像系统可客观地比较生物自发超弱发光,为进一步研究超弱发光机理提供实验基础。     

大光金鱼花不同叶位叶片超微弱发光的光子计数统计

采用生物超微弱发光探测技术,并用量子光学中的光子计数分布统计方法测量植物叶片超微弱发光的结果表明,大光金鱼花幼叶和老叶的超微弱发光强度较低,光子计数分布与泊松分布基本吻合。     

水葫芦及菠菜光合作用原初光反应的超快光谱研究

采用相同的分离技术,从水葫芦(Eichhornia crassipes(Mart)Solms.)和菠菜(Spinacia oleracea L.)叶片中提取叶绿体.利用吸收光谱和低温荧光光谱及皮秒荧光单光子计数技术对它们的光谱性质和光系统Ⅱ荧光寿命进行了研究.这两种叶绿体吸收光谱相似,暗示着它们都能高效吸收不同波长的光子.低温荧光光谱显示,水葫芦叶绿体两个光系统之间激发能分配平衡状态差,表明不利于该植物叶绿体高效利用吸收的光子能.采用三指数动力学模型对测定的光系统Ⅱ荧光衰减曲线拟合,水葫芦叶绿体光系统Ⅱ荧光衰减寿命分别是:138,521和1 494 ps;菠菜叶绿体荧光寿命分别是:197,465和1 459ps.并且归属了荧光组分,慢速度荧光衰减是由叶绿素堆积造成的,中等速度荧光衰减源于PSⅡ反应中心重新结合电荷组分,快速度荧光衰减归属于PSⅡ反应中心组分.基于20ps模型计算的水葫芦和菠菜叶绿体PSⅡ反应中心激发能转能效率分别是87%和91%.该结果与转能效率为100%的观点不一致.实验结果支持PSⅡ反应中心电荷分裂20 ps时间常数模型.根据转能效率,水葫芦生长速度不大于菠菜生长速度,但是,水葫芦叶绿体中含有丰富的胡萝卜素成分,其单位质量叶绿体吸收光能大于单位质量菠菜叶绿体吸收的量.实验结果还暗示植物叶绿体体系传能高效,接近于100%.     

捕食和食物交互作用条件下根田鼠季节性波动种群攻击水平及其行为多态性分析

在捕食和附加食物交互作用条件下,以恐吓、进攻、追逐、争斗及回避5类行为为变量,以恐吓、进攻、追逐计数之和的平均值作为攻击水平,测定根田鼠种群不同波动时期成体的攻击水平.发现根田鼠的攻击性与种群波动时期之间,存在明显的关联.统计分析结果表明,在种群3个波动时期,4种处理种群两性攻击型个体比例差异显著.除预防捕食者无附加食物（-P,-F）种群的雌体外,其它处理种群增长期和高峰期雌性和雄性攻击型个体的比例高于其衰减期.其中,预防捕食者附加食物（-P,＋F）种群、-P,-F种群及未预防捕食者附加食物（＋P,＋F）种群,雄性攻击型个体的比例均为增长期＞高峰期＞衰减期;在未预防捕食者无附加食物（＋P,-F）种群,雄性攻击型个体比例为高峰期＞增长期＞衰减期.各处理种群雌性攻击型个体比例的格局与雄体的不同.其中,-P,＋F种群及＋P,＋F种群为增长期＞高峰期＞衰减期,＋P,-F种群为高峰期＞增长期＞衰减期,而-P,-F种群攻击型个体比例为高峰期＞衰减期＞增长期.虽不同处理种群雌体及雄体的5类行为变量与种群密度的相关性不一致,而具有明显攻击性的恐吓、进攻及争斗3类行为则分别与种群密度呈显著或极显著的线性正相关关系,其结果与Chitty多态行为假设预测的一致;验证了所提出的特定假设：种群外部因子捕食和食物交互效应介导的攻击行为选择,是引起田鼠类种群季节性波动的主要内部因子.     

第四届国际光电子与微电子技术及应用大会（OMTA2020）

正2020年10月20-22日江苏南京https://b2b.csoe.org.cn/meeting/OMTA2020.html光电子与微电子技术是支撑通信网络、高性能计算、智能终端、生物医疗等应用领域的重要基础,具有广泛的应用前景和很强的研发需求。本会议将邀请100余位顶尖技术专家汇聚一堂,共同探讨光电子与微电子技术在光通信、先进传感、发光与显示、光存储、生物光电子、能源光电子等领域的最新进展,并设立拍赫兹通信和碳     

光子嫩肤用滤光片的设计与制备

分析了强脉冲光子嫩肤技术所用滤光片的特点。采用解析法构造初始膜系,单纯形法和模拟退火法相结合的优化方法设计了某型光子嫩肤仪用带通滤光片,并用盟特卡罗法分析了随机误差对滤光片光谱特性的影响,得到了理想的设计结果。采用双离子束溅射的方法制备了带通滤光片,从实测光谱图可见,通带平均透过率在95％以上,截止带平均透过率低于0．5％。各项参数均满足设计要求。     

基因工程菌在活性污泥中的衰减及其影响因素

在间歇实验和连续运行的反应器中,考察了基因工程菌细胞密度在活性污泥中的衰减,进行了动力学模型拟合,求得模型参数,并分析了影响衰减的因素.结果表明,在间歇实验和连续运行反应器中,细胞密度在活性污泥中均呈现初期快速衰减而后趋于稳定的趋势.其衰减动力学可以用Logistic模型和Gompertz模型拟合,两个模型的拟合都有很好的相关性和显著性,拟合优度没有差异.在不同的条件下,间歇实验拟合得到的参数取值范围是：Logistic模型衰减系数r′=0.5～0.6 h^-1,稳定细胞密度K′=10⁴～10⁵ cfu·ml^-1;Gompertz模型初始衰减系数b=0.6～1.2 h^-1,比衰减系数a=0.02～0.09 h^-1.初始投加密度对衰减速率影响不大,但是显著影响稳定细胞密度.污泥浓度和温度对衰减速率和稳定密度均有显著影响. 污泥浓度越大,衰减越快,稳定密度越大;而温度越低,衰减越快,稳定密度也会越低.营养条件和微型动物捕食也是影响衰减的重要因素.基因工程菌在连续运行的膜生物反应器和活性污泥反应器中的细胞密度衰减,也可以用Logistic模型拟合,其参数在间歇实验的取值范围内.     

Measurements of fluorescence lifetimes by use of a hybrid time-correlated and multifrequency phase fluorometer

Measurements of homogeneous and heterogeneous fluorescence intensity decays using a hybrid time-correlated single photon counting/multifrequency phase fluorometer are reported. A trio of fluorophores exhibiting a range of decay profiles was selected. p-Terphenyl, 1,4-bis[2-(4-methyl-5-phenyloxazolyl)]benzene [(Me)2POPOP], and p-bis[2-(5-phenyloxazolyl)]benzene (POPOP), commonly used reference fluorophores, were analyzed initially; their emissions were characterized by monoexponential decay functions. Additionally, emissions from two single tryptophan proteins with different decay profiles were measured. Scorpion neurotoxin variant 3 required three exponentials to fit the emission decay properly (average lifetime approximately 500 ps). At pH 5.5, the fluorescence emission of ribonuclease T1 showed a monoexponential decay with a measured lifetime of approximately 4.0 ns. Thus, in each case, the results from both measurements were consistent between the two detection systems, confirming the view that the two approaches for measuring fluorescence lifetimes are equivalent.     

A time-resolved fluorescence study of human copper-zinc superoxide dismutase

The intrinsic fluorescence decay of human Cu,Zn superoxide dismutase was measured by frequency-domain techniques. The protein consists of two subunits, each containing one tryptophan and no tyrosine residues. Using a synchrotron radiation source, which allows facile selection of the excitation wavelength, the dependence of the emission decay upon excitation was studied. No significant excitation wavelength effects were found. The two tryptophans contained in the dimer, although fully equivalent and exposed to solvent, showed a fluorescence decay that cannot be described by a single lifetime. Either two lifetimes, or one Lorentzian-shaped continuous distribution of lifetimes, are needed to obtain a good fit. Under identical experimental conditions, control experiments showed that N-acetyltryptophanamide, an analogue of tryptophanyl residues in proteins, decays with a single lifetime. The heterogeneous decay of tryptophan fluorescence in superoxide dismutase is interpreted as due to the presence of static and/or dynamic conformers in the protein that decay with different lifetimes. The two models of discrete lifetimes and continuous distribution of lifetimes are discussed with reference to measurements on holo- and apo-human superoxide dismutase.     

水在菌紫质光循环和质子泵中的作用

用圆二色仪和闪光动力学先谱仪分别测量了空气干燥紫膜薄层的圆二色谱及紫膜LB膜的M_(412)的衰减过程.在于燥紫膜的圆二色谱上出现412nm的正峰,它是光循环中间体M_(412)的特征峰.在无水介质中,紫膜LB膜中的BR仍能进行先化学循环而检测到中间体M_(412),但M_(412)的衰减速度减慢,产生M_(412)的堆积,质子化过程受阻.在有水的介质中,只要有足够的H~+存在,紫膜LB膜中的BR的中间体M_(412)的衰减速度明显加快.说明水介质的H~+是完成正常光化学循和质子化过程必不可少的.     

Time-resolved fluorescence and anisotropy decay of the tryptophan in adrenocorticotropin-(1-24)

The direct time-resolved fluorescence anisotropy of the single tryptophan residue in the polypeptide hormone adrenocorticotropin-(1-24) (ACTH) and the fluorescence decay kinetics of this residue (Trp-9) are reported. Two rotational correlation times are observed. One, occurring on the subnanosecond time scale, reflects the rotation of the indole ring, and the other, which extends into the nanosecond range, is dominated by the complex motions of the polypeptide chain. The fluorescence lifetimes of the single tryptophan in glucagon (Trp-25) and the 23-26 glucagon peptide were also measured. In all cases the fluorescence kinetics were satisfied by a double-exponential decay law. The fluorescence lifetimes of several tryptophan and indole derivatives and two tryptophan dipeptides were examined in order to interpret the kinetics. In close agreement with the findings of Szabo and Rayner [Szabo, A. G., & Rayner, D. M. (1980) J. Am. Chem. Soc. 102, 554-563], the tryptophan zwitterion exhibits emission wavelength dependent double-exponential decay kinetics. At 320 nm tau 1 = 3.2 ns and tau 2 = 0.8 ns, with alpha 1 = 0.7 and alpha 2 = 0.3. Above 380 nm only the 3.2-ns component is observed. By contrast the neutral derivative N-acetyltryptophanamide has a single exponential decay of 3.0 ns. The multiexponential decay kinetics of the polypeptides are discussed in terms of flexibility of the polypeptide chain and neighboring side-chain interactions.     

Constrained analysis of fluorescence anisotropy decay:application to experimental protein dynamics

Hydrodynamic properties as well as structural dynamics of proteins can be investigated by the well-established experimental method of fluorescence anisotropy decay. Successful use of this method depends on determination of the correct kinetic model, the extent of cross-correlation between parameters in the fitting function, and differences between the timescales of the depolarizing motions and the fluorophore's fluorescence lifetime. We have tested the utility of an independently measured steady-state anisotropy value as a constraint during data analysis to reduce parameter cross correlation and to increase the timescales over which anisotropy decay parameters can be recovered accurately for two calcium-binding proteins. Mutant rat F102W parvalbumin was used as a model system because its single tryptophan residue exhibits monoexponential fluorescence intensity and anisotropy decay kinetics. Cod parvalbumin, a protein with a single tryptophan residue that exhibits multiexponential fluorescence decay kinetics, was also examined as a more complex model. Anisotropy decays were measured for both proteins as a function of solution viscosity to vary hydrodynamic parameters. The use of the steady-state anisotropy as a constraint significantly improved the precision and accuracy of recovered parameters for both proteins, particularly for viscosities at which the protein's rotational correlation time was much longer than the fluorescence lifetime. Thus, basic hydrodynamic properties of larger biomolecules can now be determined with more precision and accuracy by fluorescence anisotropy decay.     

Sodium-calcium exchange affects local calcium signal decay and the rate of exocytotic secretion in single chromaffin cells

The effects of Na+ deprivation on local calcium signal decay and the rate of exocytotic secretion were measured in single bovine chromaffin cells to determine whether Na-Ca exchange influences the local cytosolic Ca2+ signal for neurohormone release. Na+ replacement with N-methylglucamine caused a marked slowing of the decay of the local Ca2+ signal near points of its initiation, as measured by high-resolution fluorescent Ca2+ imaging in the confocal laser scanning microscope. Na+ replacement also resulted in a doubling of the rate and magnitude of exocytotic secretion measured in single cells by high-resolution microamperometry. Release rates provide an independent measure of local active zone Ca2+. Five repetitive stimulations of the same cell in Na+-free, but not in Na+-containing, medium resulted in a progressively increasing rate of catecholamine release, suggesting an increasing level of active zone Ca2+ and a role of Na-Ca exchange activity in Ca2+ clearance between stimulations. As secretory activity and its triggering Ca2+ signals are known to be co-localized in active zones along the plasma membrane, the results suggest that Na-Ca exchange may influence the decay of the local Ca2+ signal for exocytotic secretion. This would be consistent with a contribution to local Ca2+ clearance by a novel mechanism utilizing the insertion of secretory vesicle Na-Ca exchangers into the plasma membrane during exocytosis.     

Fluorescence Relaxation Kinetics from Rhodopsin and Isorhodopsin

The fluorescence kinetics of bovine rhodopsin and isorhodopsin excited with a single picosecond laser pulse have been measured with a streak camera. The rise and the decay time of the intrinsic fluorescence emission from rhodopsin and isorhodopsin are found to be <12 ps.     

Application of a reference convolution method to tryptophan fluorescence in proteins. A refined description of rotational dynamics

A reference method for the deconvolution of polarized fluorescence decay data is described. Fluorescence lifetime determinations for p-terphenyl, p-bis[2-(5-phenyloxazolyl)]benzene and N-acetyltryptophanamide (AcTrpNH2) show that with this method more reliable fits of the decays can be made than with the scatterer method, which is most frequently used. Analysis of the AcTrpNH2 decay with p-terphenyl as the reference compound yields an excellent fit with lifetimes of 2.985 ns for AcTrpNH2 and 1.099 ns for p-terphenyl (20 degrees C), whereas the AcTrpNH2 decay cannot be satisfactorily fitted when the scatterer method is used. The frequency of the detected photons is varied to determine the conditions where pulse pile-up starts to affect the measured decays. At detection frequencies of 5 kHz and 15 kHz, which corresponds to 1.7% and 5% respectively of the rate of the excitation photons no effects are found. Decays measured at 30 kHz (10%) are distorted, indicating that pile-up effects play a role at this frequency. The fluorescence and fluorescence anisotropy decays of the tryptophan residues in the proteins human serum albumin, horse liver alcohol dehydrogenase and lysozyme have been reanalysed with the reference method. The single tryptophan residue of the albumin is shown to be characterized by a triple-exponential fluorescence decay. The anisotropy decay of albumin was found to be mono-exponential with a rotational correlation time of 26 ns (20 degrees C). The alcohol dehydrogenase has two different tryptophan residues to which single lifetimes are assigned. It is found that the rotational correlation time for the dehydrogenase changes with excitation wavelength (33 ns for lambda ex = 295 nm and 36 ns for lambda ex = 300 nm at 20 degrees C), indicating a nonspherical protein molecule. Lysozyme has six tryptophan residues, which give rise to a triple-exponential fluorescence decay. A single-exponential decay with a rotational correlation time of 3.8 ns is found for the anisotropy. This correlation time is significantly shorter than that arising from the overall rotation and probably originates from intramolecular, segmental motion.     

High-throughput screening in two dimensions: Binding intensity and off-rate on a peptide microarray

We report a high-throughput two-dimensional microarray-based screen, incorporating both target binding intensity and off-rate, which can be used to analyze thousands of compounds in a single binding assay. Relative binding intensities and time-resolved dissociation are measured for labeled tumor necrosis factor alpha (TNF-α) bound to a peptide microarray. The time-resolved dissociation is fitted to a one-component exponential decay model, from which relative dissociation rates are determined for all peptides with binding intensities above background. We show that most peptides with the slowest off-rates on the microarray also have the slowest off-rates when measured by surface plasmon resonance (SPR).