Investigation of the intramembrane distribution of chlorin e6 derivatives by using fluorescence quenching

Belarusian State University, 4, F. Skorina Ave., Minsk, 220050, Belarus. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 62, No. 2, pp. 179–187, March–April, 1995     

Fluorescence lifetimes of diphenylhexatriene-containing probes reflect local probe concentrations: Application to the measurement of membrane fusion

An important process in the life of a cell is fusion between cellular membranes. This is the process by which two cellular compartments surrounded by different membranes join to become a single compartment surrounded by a single membrane, without significant loss of compartment contents. To demonstrate fusion, the cell biophysicist must demonstrate all three critical aspects of the process: (1) mixing of membrane components, (2) mixing of compartment contents; and (3) retention of compartment contents. Most commonly, accomplishing this involves the use of fluorescence probes. The general theme to the methods described involves some form of concentration-dependent quenching. An unique method developed in our laboratory utilizes the concentration dependence of the fluorescence lifetime of a phosphatidylcholine containing carboxyethyl diphenylhexatriene at position 2 and palmitic acid at position 1 of glycerol (DPHpPC). The fluorescence lifetime of this molecule and that of its parent fluorophore diphenylhexatriene (DPH) shorten dramatically as their two-dimensional concentrations in a membrane increase. This lifetime quenching can be described by dimer formation that reduces the symmetry of the DPH excited state. This phenomenon allows one to use the fluorescence lifetime to gain insight into the local concentration of probe in microscopic regions of a membrane. One application of this is in distinguishing lipid transfer between the outer leaflets of two contacting membrane bilayers from fusion between these membranes that leads to mixing of lipids in both the inner and outer leaflets of the membrane bilayers. This allows a single measurement to demonstrate fusion between membrane pairs.Abbreviations PEG poly(ethylene glycol) - Na₂EDTA ethyiene-diamine-tetraacedic acid, disodium salt - LUV large, unilamellar vesicles made by rapid extrusion technique - DPH 1,6-diphenyl-trans-1,3,5-hexatriene - DPHpPC 1-palmitoyl-2-[[[2-[4- (phenyl-trans-1,3,5-hexatrienyl)phenyl]ethyl]oxy]carbonyl]-3-sn-phosphatidylcholine - DPPC 1,2-dipalmitoyl-3-sn-phosphatidylcholine - PA palmitic acid - NBD-PE N-(7-nitro-2,1,3-benzoxadiazol-4-yl)-PE - Rh-PE N-(lissamine Rhodamine B sulfoyl)-PE - R₁₈ octadecyl Rhodamine B chloride - ANTS 1-aminonaphthalene-3,6,8-trisulfonic acid - DPX N,N-p-xylylene-bis(pyradinium bromide)     

消癌平诱导人肺腺癌(ASTC-a-1)细胞内caspase-3活化的荧光光谱分析

采用CCK-8技术检测发现传统中药消癌平（XAP）对人肺腺癌（ASTC-a-1）细胞的增殖活性具有显著的抑制作用。利用共聚焦扫描荧光显微成像、荧光共振能量转移（FRET）及其受体光漂白技术证实了基于FRET技术构建的SCAT3质粒在ASTC-a-1细胞中的稳定表达。将消癌平加入细胞培养液中培育细胞,并在不同的时间检测活细胞内SCAT3的荧光发射光谱,从而监测细胞中caspase-3的活化状态。实验结果表明：(1)消癌平可以有效抑制人肺腺癌（ASTC-a-1）细胞的增殖活性并诱导细胞的死亡,消癌平对细胞的抑制作用具有剂量依赖性。(2)消癌平处理细胞72 h后,细胞内大量的SCAT3被切割,表明细胞内caspase-3的活化水平明显升高。(3)将含消癌平的细胞培养液与细胞共同培养24 h,然后采用没有消癌平的细胞培养液培养细胞48和72 h后,细胞内SCAT3的光谱没有明显改变,表明消癌平作用细胞24 h内没有显著激活细胞内的caspase-3。     

Er：YbGG纳米粉体制备及荧光发光性能研究

采用溶胶-凝胶法制备Er：YbGG纳米粉体,利用XRD、TG-DTA、IR和SEM等测试手段分析了Er：YbGG纳米粉体的物相结构和光谱性能。 结果表明:Er：YbGG纳米粉体属于立方晶系,空间群为Ia-3d,晶格参数a=1.2162 nm。在980 nm激光激发下,Er：YbGG纳米粉体在1 533 nm附近获得了较强的发射,其较高的发射强度得益于Yb³⁺-Er³⁺离子之间的能量传递。     

Effect of Calcium Ion Concentration on Small Molecule Desorption from Alginate Beads

Spherical alginate beads were prepared by ionotropic gelation of sodium alginate through the use of calcium ions. Pyranine (Py) was added to the alginate solution as a small molecule probe for fluorescence studies. Desorption of Py in water from the alginate beads cross-linked with calcium ions was studied by using the steady state fluorescence technique. The fluorescence emission intensity (I) from Py was monitored during the desorption process at 512 nm using the time drive mode of the spectrofluorometer. The increase in I was attributed to Py release from the beads. The Fickian diffusion model was used to calculate the desorption coefficients, D_, which were found to be increased up to 3% (w/v) CaCl₂ concentration in the beads, and then decreased with a further increase of CaCl₂ content. On the other hand, the encapsulation efficiency of Py in the calcium alginate beads presented the reverse behavior compared to D. It was observed that, when the content of CaCl₂ was increased, the incubation time, t_0, for the start of desorption increased.     

Back focal plane imaging of Tamm plasmons and their coupled emission

The unique optical properties of Tamm plasmons (TPs) – such as flexible wavevector matching conditions including inplane wavevector within the light line, and existing both S‐ and P‐polarized TPs − facilitate them for direct optical excitation. The Tamm plasmon‐coupled emission (TPCE) from a combined photonic–plasmonic structure sustaining both surface plasmons (SPs) and TPs is described in this paper. The sensitivity of TPCE to the emission wavelength and polarization is examined with back focal plane imaging and verified with the numerical calculations. The results reveal that the excited probe can couple with both TPs and SPs, resulting in surface plasmon‐coupled emission (SPCE) and TPCE, respectively. The TPCE angle is strongly dependent on the wavelength allowing for spectral resolution using different observation angles. These Tamm structures provide a new tool to control the optical emission from dye molecules and have many potential applications in fluorescence‐based sensing and imaging.     

多层纳米Au/DPO薄膜的荧光特性

在低压直流溅射沉积的纳米Au薄膜表面喷涂有机固体晶体2,5-二苯基恶唑（DPO）,制成具有（Au+DPO）单元结构的多层纳米薄膜。利用XRD表征多层纳米薄膜的晶体结构,通过SEM表征各层薄膜的表面形貌,并测试了多层纳米薄膜的紫外荧光特性。与纯DPO薄膜相比,多层纳米薄膜的紫外荧光峰出现了5 nm的蓝移,而且DPO荧光峰形貌发生改变。在多层膜中,DPO薄膜的荧光强度与薄膜层数成反比关系,而纳米Au膜的荧光强度与薄膜层数成正比关系。SEM与XRD测试结果表明,多层薄膜中的DPO薄膜随着薄膜层数的增加逐渐成为无定型结构。DPO薄膜与纳米Au膜相互作用,导致其晶体结构异于单层薄膜,进而改变了其荧光特性。     

基于叶绿素荧光光谱分析的稻瘟病害预测模型

为了实现稻瘟病的快速、准确和无损检测,力求构建稻瘟病害预测模型。根据水稻叶片相对病害面积将稻瘟病划分为3个等级,通过激光诱导法采集不同病害等级的活体水稻叶片叶绿素荧光光谱。选取502～830 nm波段激光诱导叶绿素荧光光谱(LICF)作为研究对象,利用Savitzky-Golay平滑法(SG)和一阶导数变换(FDT)对光谱信息进行预处理,通过主成分分析(PCA)方法获取经SG-FDT预处理后光谱的特征向量,根据累积贡献率和方差选取前3个主成分进行分析。将试验样本分为建模样本和检验样本,以稻瘟病害等级为预测指标,利用建模样本的133片叶片的光谱和病害信息分别结合判别分析(DA)、多类逻辑回归分析(MLRA)和多层感知器(MLP)建立稻瘟病的预测模型,利用检验样本的89片叶片的光谱和病害信息对所建模型进行预测检验,完成对PCA-DA、PCA-MLRA和PCA-MLP的对比寻优。结果表明,PCA-DA,PCA-MLRA和PCA-MLP模型均能完成对稻瘟病害的预测,但PCA-MLP模型的平均预测准确率能够达到91.7%,相比PCA-DA和PCA-MLRA模型,在稻瘟病害3个等级上均具有较好的分类和预测能力。     

Recent advances in rice genome and chromosome structure research by fluorescence in situ hybridization (FISH)

Fluorescence in situ hybridization (FISH) is an effective method for the physical mapping of genes and repetitive DNA sequences on chromosomes. Physical mapping of unique nucleotide sequences on specific rice chromosome regions was performed using a combination of chromosome identification and highly sensitive FISH. Increases in the detection sensitivity of smaller DNA sequences and improvements in spatial resolution have ushered in a new phase in FISH technology. Thus, it is now possible to perform in situ hybridization on somatic chromosomes, pachytene chromosomes, and even on extended DNA fibers (EDFs). Pachytene-FISH allows the integration of genetic linkage maps and quantitative chromosome maps. Visualization methods using FISH can reveal the spatial organization of the centromere, heterochromatin/euchromatin, and the terminal structures of rice chromosomes. Furthermore, EDF-FISH and the DNA combing technique can resolve a spatial distance of 1 kb between adjacent DNA sequences, and the detection of even a 300-bp target is now feasible. The copy numbers of various repetitive sequences and the sizes of various DNA molecules were quantitatively measured using the molecular combing technique. This review describes the significance of these advances in molecular cytology in rice and discusses future applications in plant studies using visualization techniques.