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1.
The labeling strategy with gold nanoparticles for the conventional surface plasmon resonance (SPR) signal enhancement has been frequently used for the sensitive determination of small molecules binding to its interaction partners. However, the influence of gold nanoparticles with different size and shape on SPR signal is not known. In this paper, three kinds of gold nanoparticles, namely nanorods, nanospheres, and nanooctahedrons with different size, were prepared and used to investigate their effects on the conventional SPR signal at a fixed excitation wavelength 670 nm. It was found that the SPR signal (i.e., resonant angle shift) was varied with the shapes and sizes of gold nanoparticles in suspension at a fixed concentration due to their different plasmon absorbance bands. For gold nanorods with different longitudinal absorbance bands, three conventional SPR signal regions could be clearly observed when the gold nanorod suspensions were separately introduced onto the SPR sensor chip surface. One region was the longitudinal absorbance bands coinciding with or close to the SPR excitation wavelength that suppressed the SPR angle shift. The second region was the longitudinal absorbance bands at 624 to 639 and 728 to 763 nm that produced a moderate increase on the SPR resonant angle shift. The third region was found for the longitudinal absorbance bands from 700 to 726 nm that resulted in a remarkable increase in the SPR angle shift responses. This phenomenon can be explained on the basis of calculation of the correlation of SPR angle shift response with the gold nanorod longitudinal absorbance bands. For nanospheres and nanooctahedrons, the SPR angle shift responses were found to be particle shape and size dependent in a simple way with a sustaining increase when the sizes of the nanoparticles were increased. Consequently, a guideline for choosing gold nanoparticles as tags is suggested for the SPR determination of small molecules with binding to the immobilized interaction partners.  相似文献   

2.
A tunable fourth derivative UV absorbance method based on a variable spectral shift has been developed and compared to the Savitzky-Golay method and the analytical derivative. The parameters of the method were optimised for the analysis of the UV absorbance spectra of the aromatic amino acids to quantify the effect of decreasing solvent polarity on their fourth derivative spectra. The wavelength of the highest maximum (max) (for tyrosine and phenylalanine) or the amplitude of the highest maximum (Amax) (for tryptophan), were shown to depend linearly on the dielectric constant of the solvent, ranging from water to cyclohexane. The only effect of pressure in the 1 to 500 MPa range is a small decrease in the fourth derivative amplitude. This method appears therefore as a suitable tool to evaluate changes of the dielectric constant in the vicinity of the aromatic amino acids in proteins which undergo pressure induced structural changes.  相似文献   

3.
The ultraviolet (UV) spectra of selected enaminones were determined in acidic, alkaline and neutral media and compared to their anticonvulsant activities. The wavelength of maximum absorption and molar absorptivity were compared with the anticonvulsant activity of the selected secondary and tertiary enaminones, and general inferences were made. The UV spectra of the enaminones had hypsochromic shifts in acidic media in comparison with neutral media. Generally, a small hypsochromic shift occurred in alkaline media when compared to the neutral solutions of the enaminones. The tertiary enaminones absorbed UV light at longer wavelength than the secondary enaminones in acidic, neutral and alkaline media. In particular, the tertiary enaminones displayed absorption at the higher end and secondary enaminones towards the lower end of the UV wavelength range 292-315nm in aqueous media. Tertiary enaminones (30-33) which were devoid of the NH proton were found to be uniformly inactive in a mouse model of electroshock seizures, while some secondary enaminones (1, 5-8, 12, 16, 18, 20, 23-25, 28 and 29) had anticonvulsant activity. Thus the NH group of secondary enaminones is very important for anticonvulsant activity, and this agrees with an already established trend in proton NMR spectroscopy. In addition, the para-substitution on the phenyl group in some enaminones result in higher molar absorptivity (epsilon) values that enhance anticonvulsant activity. These results indicate that the anticonvulsant activity of enaminones is not due to electronic effect alone, but is probably due to a combination of factors including electronic and steric effects, lipophilicity, and hydrogen bonding.  相似文献   

4.
Zebrafish and goldfish are both diurnal freshwater fish species belonging to the same family, Cyprinidae, but their visual ecological surroundings considerably differ. Zebrafish are surface swimmers in conditions of broad and shortwave-dominated background spectra and goldfish are generalized swimmers whose light environment extends to a depth of elevated short wavelength absorbance with turbidity. The peak absorption spectrum (lambdamax) of the zebrafish blue (SWS2) visual pigment is consistently shifted to short wavelength (416 nm) compared with that of the goldfish SWS2 (443 nm). Among the amino acid differences between the two pigments, only one (alanine in zebrafish and serine in goldfish at residue 94) was previously known to cause a difference in absorption spectrum (14-nm lambdamax shift in newt SWS2). In this study, we reconstructed the ancestral SWS2 pigment of the two species by applying likelihood-based Bayesian statistics and performing site-directed mutagenesis. The reconstituted ancestral photopigment had a lambdamax of 430 nm, indicating that zebrafish and goldfish achieved short wavelength (-14 nm) and long wavelength (+13 nm) spectral shifts, respectively, from the ancestor. Unexpectedly, the S94A mutation resulted in only a -3-nm spectral shift when introduced into the goldfish SWS2 pigment. Nearly half of the long wavelength shift toward the goldfish pigment was achieved instead by T116L (6 nm). The S295C mutation toward zebrafish SWS2 contributed to creating a ridge of absorbance around 400 nm and broadening its spectral sensitivity in the short wavelength direction. These results indicate that the evolutionary engineering approach is very effective in deciphering the process of functional divergence of visual pigments.  相似文献   

5.
Knoevenagel condensation of phenothiazine-3,7-dicarbaldehyde with an isophorone yielded a new phenothiazine derivative ( PTZ-c ) fluorophore. The solvatochromic and pH-sensing abilities of PTZ-c , an asymmetric fluorophore with a single isophorone molecule, were shown to be exceptional. PTZ-c produced very delicate absorbance and emission spectra. When the polarity of the solvent was increased, the PTZ-c emission spectra showed greater sensitivity than the absorption spectra. Multiple spectroscopic techniques, including Fourier transform infrared spectroscopy, nuclear magnetic resonance, and mass spectrometry, were used to characterize the manufactured PTZ-c sensor. To demonstrate the beneficial solvatochromic behaviour associated with intramolecular charge transfer, the absorption spectra of the synthesized DA PTZ-c dye were analyzed in different solvents of varying polarity. Band intensity and the wavelength of PTZ-c emission were also found to be highly solvent dependent. It was observed that when solvent polarity was increased to a maximum of 4122 cm−1, Stokes’ shift also increased. To analyze the Stokes' shift that depended on the solvent, a linear correlation between solvation and energy was used. An investigation of PTZ-c quantum yield (ф) was also conducted. Both the absorbance and fluorescence spectra of the sensor in dimethylformamide as a function of pH were studied. A fluorescence peak was seen at 562 nm, whereas the greatest absorption wavelengths were found at 403 and 317 nm. It was shown that the pH-sensing mechanism depended on protons removed from the PTZ-c chromophore, which caused a colour shift and variation in both emission and colorimetric properties.  相似文献   

6.
The adaptation of a commercially available dual wavelength/split beam spectrophotometer for the real time acquisition by a minicomputer of absorbance spectra is described. The computer software and interface are designed so that: a) The normal operation of the spectrophotometer is unaltered; b) the adaptation of the spectrophotometer for computer acquisition of spectra is relatively simple and only a rudimentary knowledge of electronics and computer programming is required.The utility of this adaptation of a dual wavelength/split beam spectrophotometer for baseline correction, enhancement of the sensitivity of the spectrophotometer and subsequent analysis of recorded absorbance spectra is described.  相似文献   

7.
A novel method for sensitivity enhancement of spectral surface plasmon resonance (SPR) biosensors was presented by reducing the refractive index of the sensing prism in the analysis of protein arrays. Sensitivity of spectral SPR biosensors with two different prisms (BK-7, fused silica) was analyzed by net shifts of resonance wavelength for specific interactions of GST–GTPase binding domain of p21-activated kinase-1 and anti-GST on a mixed thiol surface. Sensitivity was modulated by the refractive index of the sensing prism of the spectral SPR biosensors with the same incidence angle. The sensitivity of a spectral SPR biosensor with a fused silica prism was 1.6 times higher than that with a BK-7 prism at the same incidence angle of 46.2°. This result was interpreted by increment of the penetration depth correlated with evanescent field intensity at the metal/dielectric interface. Therefore, it is suggested that sensitivity enhancement is readily achieved by reducing the refractive index of the sensing prism of spectral SPR biosensors to be operated at long wavelength ranges for the analysis of protein arrays.  相似文献   

8.
Carcinoembryonic antigen (CEA) was used as a separator to prevent the Rhodamine 6G (R6G)-induced aggregation of colloidal gold nanoparticles. The destroyed aggregation has been monitored by measuring the absorption and resonance light scattering peaks corresponding to the longitudinal surface plasmon resonance (SPR) of the chain-like aggregated gold nanoparticles (AuNPs). It was found that the pre-adding of CEA with different concentrations to the gold colloids before mixing them with R6G could lead to the longitudinal SPR peak decrease and blue shift. By analysing the intensity changing and wavelength shifting of the absorption spectra, CEA could be detected in a linear range from 0.2 to 4 ng/mL, and the limit of detection reaches to 0.1 ng/mL. The sensitivity of the CEA concentration dependent shifting and quenching of the plasmonic absorption and scattering corresponding to the AuNPs aggregation presents a well potential application of biologic spectral sensing.  相似文献   

9.
The squirrel monkey (Saimiri sciureus) exhibits a polymorphism of colour vision: some animals are dichromatic, some trichromatic, and within each of these classes there are subtypes that resemble the protan and deutan variants of human colour vision. For each of ten individual monkeys we have obtained (i) behavioural measurements of colour vision and (ii) microspectrophotometric measurements of retinal photopigments. The behavioural tests, carried out in Santa Barbara, included wavelength discrimination, Rayleigh matches, and increment sensitivity at 540 and 640 nm. The microspectrophotometric measurements were made in London, using samples of fresh retinal tissue and a modified Liebman microspectrophotometer: the absorbance spectra for single retinal cells were obtained by passing a monochromatic measuring beam through the outer segments of individual rods and cones. The two types of data, behavioural and microspectrophotometric, were obtained independently and were handed to a third party before being interchanged between experimenters. From all ten animals, a rod pigment was recorded with lambda max (wavelength of peak absorbance) close to 500 nm. In several animals, receptors were found that contained a short-wave pigment (mean lambda max = 433.5 nm): these violet-sensitive receptors were rare, as in man and other primate species. In the middle- to long-wave part of the spectrum, there appear to be at least three possible Saimiri photopigments (with lambda max values at about 537,550 and 565 nm) and individual animals draw either one or two pigments from this set, giving dichromatic or trichromatic colour vision. Thus, those animals that behaviourally resembled human protanopes exhibited only one pigment in the red-green range, with lambda max = 537 nm; other behaviourally dichromatic animals had single pigments lying at longer wavelengths and these were the animals that behaviourally had higher sensitivity to long wavelengths. Four of the monkeys were behaviourally judged to be trichromatic. None of the latter animals exhibited the two widely separated pigments (close to 535 and 567 nm) that are found in the middle- and long-wave cones of macaque monkeys.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

10.
Microspectrophotometric measurements were performed on 217 photoreceptors from cynomolgus, Macaca fascicularis, and rhesus, M. mulatta, monkeys. The distributions of cell types, for rods and blue, green, and red cones were: 52, 12, 47, and 44, respectively, for the cynomolgus, and 22, 4, 13, and 13 for the rhesus. Visual cells were obtained fresh (unfixed), mounted in various media (some containing 11-cis-retinal), and then located visually under dim red (650 nm) illumination. Absolute absorbance (A), linear dichroism (LD), and bleaching difference (BD) absorbance spectra were recorded through the sides of outer segments. The spectra were subjected to rigorous selection criteria, followed by digital averaging and Fourier transform filtering. Statistical methods were also applied to the accepted samples in the estimation of population means and variances. The wavelength of mean peak absorbance (lambda max) and the standard error at 95% certainty of the rod and blue, green, and red cone pigments in cynomolgus were 499.7 +/- 2.5, 431.4 +/- 4.2, 533.9 +/- 2.4, and 565.9 +/- 2.8 nm, respectively. The rhesus pigments were statistically indistinguishable from the cynomolgus, having lambda max of approximately 500, 431, 534, and 566 nm. Statistical tests did not reveal the presence of a lambda max subpopulation (i.e., anomalous pigments). The bandwidth of each alpha-band was determined in two segments, giving rise to the longwave half-density (LWHDBW), shortwave half-density (SWHDBW), and total half-density (THDBW) bandwidths. The LWHDBW was found to have the smallest variance. Both the LWHDBW and the THDBW showed linear dependence on the peak wavenumber (lambda max)-1 for the four macaque pigments.  相似文献   

11.
用分光光度法在体外研究了不同pH条件下马铃薯'转心乌'块茎花色苷颜色呈现和降解速率的变化,以探讨马铃薯'转心乌,块茎颜色呈现的机理.结果显示:在pH 2.0时,该花色苷呈现最强烈的红色.随着pH从0增加到13.0,该花色苷在可见光区的最大吸收波长(λvis max)依次出现红移、蓝移,然后消失,在可见光区的最大吸收波长处的吸光值(Aλ,vis max)呈现为一条单峰曲线,峰值在pH 2.0处.当原始pH值被恢复到2.0后,如果原始pH小于或等于5.0,花色苷的红色均被恢复得更浓烈,λvis max不同程度地趋向537 nm,Aλ,vis max增加;如果原始pH大于或等于6.0,花色苷的红色根本不能被恢复,λvis max几乎不变,Aλ,vis max仍然维持低水平.在15℃、黑暗中,该花色苷在pH 0~5.0条件下均随时间推移而降解,在pH 2.0时的降解速度最慢;当pH小于或等于3.0时,该花色苷总体上降解缓慢,而且降解过程基本符合一级反应动力学.研究表明,'转心乌'块茎花色苷在可见光区的吸收光谱和在15℃、黑暗中的降解速度均具pH依赖性.  相似文献   

12.
A new typical phenothiazine compound functionalized with thienyl‐indandione derivative (PTZTID) was synthesized and characterized using spectral analysis (ultraviolet–visible (UV–vis) light, infrared (IR), 1H nuclear magnetic resonance (NMR) and 13C NMR tools). The UV–vis absorption spectra of the PTZTID solution in 1,4‐dioxane showed two absorption bands attributed to localized aromatic π–π* transitions of conjugated aromatic moieties and intramolecular charge transfer with the characteristics of a π–π* transition. The fluorescence spectra exhibited a maximum emission wavelength at 580 nm. The effect of concentration on photophysical properties took the form of a minor hypsochromic shift, which was attributed to some extent to the occurrence of H‐type aggregation of the PTZTID derivative. Binary solvent effects on the spectroscopic behaviour of PTZTID were measured at different H2O/1,4‐dioxane ratios. Similarly, when increasing the water content, a hypsochromic shift was observed that resulted from H‐type aggregation. Furthermore, geometry and electronic configurations of PTZTID were studied at density functional theory /B3LYP level and indicated that the compound had a nonplanar (butterfly structure).  相似文献   

13.
An absorption-based surface plasmon resonance (SPR(Abs)) biosensor probe has been developed for simple and reproducible measurements of hydrogen peroxide using a modified Trinder's reagent (a chromogenic reagent). The reagent enabled the determination of the hydrogen peroxide concentration by the development of deep color dyes (lambda(max)=630nm) through the oxidative coupling reaction with N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethylaniline sodium salt monohydrate (MAOS; C(13)H(20)NNaO(4)S.H(2)O) and 4-aminoantipyrine (4-AA) in the presence of hydrogen peroxide and horseradish peroxidase (HRP). In the present study, urea as an adduct of hydrogen peroxide for color development could be omitted from the measurement solution. The measurement solution containing 5mM hydrogen peroxide was deeply colored at a high absorbance value calculated as 46.7cm(-1) and was directly applied to the SPR(Abs) biosensing without dilution. The measurement was simply performed by dropping the measurement solution onto the surface of the SPR sensor probe, and the SPR(Abs) biosensor response to hydrogen peroxide was obtained as a reflectivity change in the SPR spectrum. After investigation of the pH profiles in the SPR(Abs) biosensor probe, a linear calibration curve was obtained between 1.0 and 50mM hydrogen peroxide (r=0.991, six points, average of relative standard deviation; 0.152%, n=3) with a detection limit of 0.5mM. To examine the applicability of this SPR(Abs) biosensor probe, 20mM glucose detection using glucose oxidase was also confirmed without influence of the refractive index in the measurement solution. Thus, the SPR(Abs) biosensor probe employing the modified Trinder's reagent demonstrated applicability to other analyte biosensing tools.  相似文献   

14.
We have investigated the sensitivity of ex situ (analysis under air condition) and in situ (analysis under liquid condition) spectral SPR sensors, which were self-constructed with fiber optic spectrometers. The sensitivity of SPR sensors was analyzed in the wavelength range of 550-780 nm by the interactions of streptavidin and biotinylated IgG, and the sensitivity was dependent on the wavelength of measurements. The sensitivity of an ex situ SPR sensor operated at the long wavelength range from 712 nm was approximately 2.6 times higher than that at the short wavelength range from 571 nm. In addition, the sensitivity of an ex situ spectral SPR sensor was about twice as high as that of an in situ spectral SPR sensor for the same resonance wavelength range. This was interpreted in that the difference in sensitivity between two SPR sensors was significantly caused by the evanescent field intensity at the metal/dielectric interface. Thus, it was suggested that ex situ spectral SPR sensors operated at the long wavelength range are sensitive biosensors for the high-throughput analysis of protein interactions on protein arrays.  相似文献   

15.
Addition of Cu2+ at low concentrations, to intact cells of the cyanobacterium, Spirulina platensis, at room temperature, caused an enhancement in intensity of fluorescence emitted by phycocyanin and induced a blue shift at the emission peak, both of which indicated changes in energy transfer within the phycobillisomes. Cu2+ also suppressed the whole-chain electron transport activity (H2O→MV) and water-splitting activity of the photosystem Ⅰ. When isolated phycocyanin and allophycocyanin were exposed to very low concentrations of Cu2+ ions, C-phycocyanin but not allophycocyanin, exhibited decrease not only in the absorbance in the longer wavelength (616--620 nm) region, but also in the fluorescence emission intensity at 647 nm accompanied by a blue shift to 643 nm. These results suggested that Cu2+ selectively bleach C-phycocyanin.  相似文献   

16.
Binding of reduced nicotinamide adenine dinucleotide to yeast alcohol dehydrogenase results in a hypsochromic shift of its absorbance maximum at 340 nm. Application of high hydrostatic pressure to the enzyme-nucleotide complex returns the absorbance maximum to longer wavelengths. This pressure-dependent bathochromic shift validates one of two assignments on the effects of pressure on the kinetics of the enzymatic oxidation of benzyl alcohol, namely the protein-ligand conformational change of the capturing form of enzyme.  相似文献   

17.
A temperature sensor based on hollow fiber (HF) filled with graphene-Ag composite nanowire and liquid is presented. The coupling properties and sensing performance are numerically analyzed by finite element method using wavelength and amplitude interrogations. Results show that the sensor exhibiting strong birefringence with x-polarized peak provides much higher sensitivities and better signal-to-noise ratio (SNR) than y-polarized, which is more suitable for temperature detection. The graphene-Ag composite nanowire can not only solve the oxidation problem but also avoid the metal coating. Moreover, it provides better performance than other similar works like Au-Ag nanowire-filled, Au nanowire-filled, and Ag nanowire-filled sensors. Contrary to the blue shift of traditional SPR temperature sensors, the resonance peak shifts to the longer wavelength in our device when temperature increases and the high sensitivity 9.44 nm/ °C is obtained. The influences of nanowire diameter, grapheme-layer thickness on the designed sensor, are also investigated. This work can provide a reference for developing a high sensitivity, real-time, remote sensing, and distributed temperature SPR sensor.  相似文献   

18.
Satoshi Hoshina 《BBA》1981,638(2):334-340
Temperature-dependent spectral changes of chlorophyll a (Chl a) incorporated into liposomes of two types of phosphatidylcholine are studied. When Chl a incorporated into the liposomes is cooled down to 5°C from the temperature of the gel-to-liquid crystalline phase transition of the lipid, the red shift as well as the increase in half-bandwidth of the red peak of Chl a are only slight. By measuring the difference spectra produced by substracting the absorption spectrum at the phase transition temperature of the lipid from that at lower temperature, it is shown that the component absorbing at longer wavelength (675–685 nm) than the peak of the red maximum (about 670 nm) significantly increases at the expense of the component absorbing at shorter wavelength (657–668 nm). The positions of positive and negative peaks depend on the temperature and the molar ratio of the lipid to Chl a. The absorbance change is most pronounced on cooling below the phase transition temperature of the lipid. The temperature-induced absorbance change is almost completely reversible. The results indicate that the aggregated forms of Chl a in liposomes can be spectrophotometrically detected in the gel phase of the lipid.  相似文献   

19.
In wavelength surface plasmon resonance (SPR) biosensor, the manipulation of SPR dispersion relation by Ag/Au bi-metallic film was first time implemented. Due to the enhanced resonant wavelength shift and the sharper SPR slope of using Ag/Au bi-metallic film, the illuminated color of reflection shows one order of magnitude greater contrast than conventional SPR biosensors. Such an Ag/Au bi-metallic film based color SPR biosensor (CSPRB) allows the detail bio-interactions, for example 100 nM streptavidin, to be distinguished by directly observing the color change of reflection through naked eyes rather than the analysis of spectrometer. In addition to the enhanced sensitivity and color contrast, this CSPRB also possesses a great linear detection range up to 0.0254 RIU, which leading to the application of point-of-care tests.  相似文献   

20.
We study an active modulation of surface plasmon resonance (SPR) of Au nanoparticles based on highly doped graphene in visible and near-infrared regions. We find that compared to the traditional metal SPR, the SPR of Au nanoparticles based on graphene causes a remarkable blue shift. The field intensity in the gap is redistributed to standing wave. The field intensity of standing wave is about one order of magnitude higher than the traditional model. Moreover, the SPR of Au nanoparticles can be actively modulated by varying the graphene Fermi energy. We find the maximum modulation of field intensity of absorption spectra is more than 21.6 % at λ?=?822?nm and the amount of blue shift is 17.4 nm, which is about 2.14 % of the initial wavelength λ 0?=?813.4?nm, with increasing monolayer graphene Fermi energy from 1.0 to 1.5 ev. We find that the SPR sensitivity to the refractive index n of the environment is about 642 nm per refractive index unit (RIU). The SPR wavelengths have a big blue shift, which is about 33 nm, with increasing number of graphene layers from 1 to 3, and some shoulders on the absorption spectra are observed in the models with multilayer graphene. Finally, we study the Au nanorod array based on monolayer graphene. We find that the blue shift caused by the graphene increases from 14 to 24 nm, with increasing gap g y from 10 to 20 nm. Then, it decreases from 24 to 14 nm, with increasing gap g y from 20 to 50 nm. This study provides a new way for actively modulating the optical and optoelectronic devices.  相似文献   

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