基于SiPM单光子探测器的荧光光谱仪研究

为了解决激光诱导荧光检测系统存在的光学结构复杂、体积大、成本高以及灵敏度不足等主要问题,研制了一种高灵敏、小型化的荧光光谱仪。该光谱仪以349 nm半导体激光器作为激发光源,采用正交型光路,将4×4窄带滤光片阵列与具有单光子灵敏度的硅光电倍增器（SiPM）阵列耦合,可实现光谱信息的多通道探测,具备结构紧凑、成本较低以及稳定性好等优点。以荧光素钠为测试样品,对光谱仪性能进行了评估。实验结果表明,光谱仪的检测限优于5×10?11 mol·L?1,在5×10?11 mol·L?1到1×10?9 mol·L?1的溶液浓度范围内,被测溶液浓度与检测所得荧光强度满足良好的线性关系,线性相关系数为0.998 39。此外,光谱仪还具备良好的重复性,荧光峰值强度的相对标准偏差小于10%。因此,该光谱仪兼具灵敏度高、线性度好、重复性与可靠性强等优点,可以满足现场实时检测的需求。     

光纤技术在近红外光谱仪中的应用

探讨了光纤在近红外光谱技术应用中光纤材料选择、能量损耗和使用注意事项等问题。通过把光纤与近红外光谱技术相结合,使得近红外光谱仪从实验室走向现场,可适用于军事恶劣、危险和复杂的环境。为在军事条件下近红外光谱技术的应用提供了更大的空间。     

X射线荧光光谱仪在PTA生产中的应用

以对二甲苯为原料 ,醋酸钴、醋酸锰为催化剂、氢溴酸为助催化剂高温氧化生产精对苯二甲酸中 ,溴以 Br-,游离态溴和有机溴化合物形式存在 ,对该氧化反应均有催化作用 ,正确测定总溴十分重要 ,常用的电位滴定法无法测定总溴的浓度。本文建立了用 X射线荧光光谱仪同时测定反应体系中 Co、Mn和总Br的方法。该法简便、可靠 ,可以在 PTA生产中使用。     

高光谱分辨率横向剪切静态干涉光谱仪

提出一种基于新型的分光方法的横向剪切分束器,分析了该分束器结构的分光原理,并对其用于干涉光谱仪中的效果进行了分析计算.该分束器同体积条件下较目前的分束器可以产生十倍以上的剪切量,应用到干涉仪中可以在获得高光谱分辨率的同时不增大仪器的体积和重量.同时,该分束器分光时,无光能返回光源,较Sagnac型分束器对光能的利用率提高了近一倍;相对变形Mach-Zehnder型结构易实现实体化.     

高光谱探测绿色涂料伪装的光谱成像研究

基于现有伪装涂料与植被反射光谱的本质差异,提出一种可有效识别当前所有绿色伪装涂料的高光谱成像方法.通过分析绿色伪装涂料与被子植物叶片的反射光谱及其一阶微分谱的差异,确定了星载和机载高光谱遥感探测中,可见光波段的绿色反射峰和780～1 300 nm的“近红外高原”波段反射率的波动性是识别绿色伪装涂料的有效光谱特征.对“近红外高原”波段的反射光谱进行成像是高光谱探测实现伪装目标可视识别的可行方法,尤其是对反射光谱一阶微分处理后进行成像可更加有效地识别植被环境中的绿色伪装涂料.     

西门子SRS300型X-荧光光谱仪故障解析

通过对西门子SRS30 0型X 荧光光谱仪几种故障原因分析以及对该仪器有关部件的维修 ,使仪器使用效率得到提高 ,同时也使其使用寿命得以延长。     

宽光谱棱镜型太阳光谱仪设计

为实现大气层外太阳光谱辐照度(SSI)变化的长期例行监测,设计了一种星载宽光谱太阳光谱仪结构。全系统仅使用单片折反式曲面棱镜实现太阳光谱250～2500 nm的分光,并通过棱镜转动实现谱平面上多探测器的同步扫描探测;同时基于Huygens子波点扩展函数（PSF）仿真了光谱仪的光谱响应函数（SRF）和光谱分辨率。分光棱镜在±2.5°扫描转角内的全谱段子午像差小于8μm;光谱分辨率在紫外谱段(250～400 nm)为0.7～3.5 nm,可见/近红外谱段(400～1000 nm)为3.5～35.0 nm,短波红外谱段(1000～2500 nm)内为28.5～41.2 nm。整个系统结构简单紧凑,性能稳定可靠,分光和像差校正能力满足大气层外太阳光谱辐照度长期监测需求。     

高分辨率傅里叶变换成像光谱仪光谱重建并行算法

高分辨率傅里叶变换成像光谱仪具有高空间分辨率和高光谱分辨率的特点,但光谱重建时间冗长。通过对傅里叶变换光谱重建流程分析,为研制的1024pixel(光谱维)×1024piexl(像宽)×1024piexl(像高)高分辨率紫外傅里叶变换成像光谱仪的数据立方体反演,设计了一种并行优化算法。实验表明,在6核处理器上对512M和2G的数据立方体进行变换,时间分别只需88.33s和489.75s,加速比分别为3.70和3.04,大幅度提高了运算效率。如将该算法应用到更多内核处理器上,可得到更高的加速比和更少的运算时间。     

Determination of photosensitizer concentration in biological tissues from diffuse reflectance and fluorescence

We present a fiber-optic device for noninvasive determination of the optical properties of biological tissues and photosensitizer concentration. The device developed can be used in two modes: the mode for detection of the spatial distribution of diffuse reflectance, and the fluorescence mode. We tested the device in vitro on specimens simulating tissue and on whole blood, and also in vivo on rats. We have shown that with additional modification and adaptation, the device can be used for in vivo monitoring of parameters important for photodynamic therapy.     

Studies on the interaction of gliclazide with bovine serum albumin by fluorescence and synchronous fluorescence spectroscopy

The interaction between gliclazide and bovine serum albumin was investigated by fluorescence and synchronous fluorescence spectroscopy. From the experimental results, it was found that the quenching mechanism was static. The results of the synchronous fluorescence obtained indicated that the binding of gliclazide with bovine serum albumin could affect conformation in bovine serum albumin. In addition, the binding constants (K_a), binding sites (n), thermodynamic parameters, binding forces, Hill’s coefficient, and binding rate of gliclazide to protein calculated from two methods using the same equation were consistent at three different temperatures (298, 310, 318 K). This indicated that as a useful supplement to the conventional method, synchronous fluorescence spectroscopy could be used to study the mechanism of drugs and proteins. The conclusion was verified by UV/vis method.     

A review on the methods for correcting the fluorescence inner-filter effect of fluorescence spectrum

Fluorescence spectroscopy is frequently used to analyze the concentration of fluorescent materials in solution. However, in conventional fluorescence spectroscopy, the response between the fluorescence intensity and fluorophore concentration is nonlinear at high concentrations due to uncompensated inner-filter effects (IFE). Many methods to resolve this problem have been developed in recent decades. This review introduces the methods used to correct the IFE, including direct correction and parameter correction. Relevant detection parameters, including the materials, matrices, detection limits, detection instruments and relative standard deviations, are tabulated. The advantages and limitations of these correction techniques are also discussed. Finally, the methods used to correct for the IFE are summarized, and future research directions are discussed.     

Review of fluorescence anisotropy decay analysis by frequency-domain fluorescence spectroscopy

This didactic paper summarizes the mathematical expressions needed for analysis of fluorescence anisotropy decays from polarized frequency-domain fluorescence data. The observed values are the phase angle difference between the polarized components of the emission and the modulated anisotropy, which is the ratio of the polarized and amplitude-modulated components of the emission. This procedure requires a separate measurement of the intensity decay of the total emission. The expressions are suitable for any number of exponential components in both the intensity decay and the anisotropy decay. The formalism is generalized for global analysis of anisotropy decays measured at different excitation wavelengths and for different intensity decay times as the result of quenching. Additionally, we describe the expressions required for associated anisotropy decays, that is, anisotropy decays where each correlation time is associated with a decay time present in the anisotropy decay. And finally, we present expressions appropriate for distributions of correlation times. This article should serve as a reference for researchers using frequency-domain fluorometry.     

Laser-induced fluorescence spectroscopy of FeS in the visible region

The laser-induced fluorescence excitation spectra of jet-cooled FeS molecules have been recorded in the energy range of 18 900-21 600 cm⁻¹, in which four parallel and one perpendicular transitions were identified for the first time. Spectroscopic constants of the observed excited states of FeS were determined by analyzing their rotationally resolved spectra. In addition, the lifetimes of most observed bands were also measured.     

Emerging biomedical and advanced applications of time-resolved fluorescence spectroscopy

Time-resolved fluorescence spectroscopy is presently regarded as a research tool in biochemistry, biophysics, and chemical physics. Advances in laser technology, the development of long-wavelength probes, and the use of lifetime-based methods, are resulting in the rapid migration of timeresolved fluorescence to the clinical chemistry lab, the patient's bedside, and even to the doctor's office and home health care. Additionally, time-resolved imaging is now a reality in fluorescence microscopy and will provide chemical imaging of a variety of intracellular analytes and/or cellular phenomena. Future horizons of state-of-the-art spectroscopy are also described. Two photon-induced fluorescence provides an increased information content to time-resolved data. Two photoninduced fluorescence, combined with fluorescence microscopy and time-resolved imaging, promises to provide detailed three-dimensional chemical imaging of cells. Additionally, it has recently been demonstrated that the pulses from modern picosecond lasers can be used to quench and/or modify the excited-state population by stimulated emission since the stimulated photons are directed along the quenching beam and are not observed. The phenomenon of light quenching should allow a new class of multipulse time-resolved fluorescence experiments, in which the excited-state population is modified by additional pulses to provide highly oriented systems.     

On the fluorescence of C_60 at room temperature

The fluorescence properties of C 60 in different organic solvents have been investigated at room temperature. Three fluorescence emission centers are discovered and ascribed to different aggregations of C 60 in solvent. A series of blue fluorescence peaks centered at 440 nm derive from C 60 nanoparticles; a distinctive yellow-green fluorescence band in 575 nm region arises from the aggregates of C 60 nanoparticles; a more informative salmon fluorescence band around 700 nm originates from C 60 microcrystals....     

Mechanistic studies of oxidation reactions by fluorescence spectroscopy: a critical assessment

The synthesis and properties of a ratiometric fluorescent dye system as a probe for the detection of oxidizing species are described. The dye consists of a strongly fluorescent Bodipy label attached to a styryl moiety. The oxidation of the exocyclic double bond by agents for syn‐ and anti‐ dihydroxylation leads to a visible change in fluorescence colour from orange to green. IR spectroscopy provides evidence for the generation of the epoxide in the reaction with peroxides, whereas a ketone and a product of unknown structure are produced in the reaction with osmiumtetroxide, each in contrast to the expected hydroxylations. Based on these first attempts, we discuss the significance of fluorescence spectroscopy to track chemical reactions at low concentrations and give a perspective of how present shortcomings can be overcome. Copyright © 2009 John Wiley & Sons, Ltd.     

Extrinsic fluorescence probe study of human serum albumin using Nile red

Nile red bound to human serum albumin (HSA) shows an order of magnitude increase in the probe's fluorescence intensity. Here, we report on the fluorescence characteristics of the probe-protein complex in Trizma buffer (pH 7.1), urea, guanidine hydrochloride, and AOT/isooctane/buffer reverse micelles using both steady—state and time-resolved fluorescence techniques. With a view to illustrating the use of extrinsic probe fluorescence spectroscopy in protein research, we demonstrate that protein unfolding can be observed through measurements of the probe's time-resolved anisotropy and steady-state fluorescence spectrum. Moreover, this shows that thermal unfolding is fundamentally different from using denaturant, with respect to changes in both the nanosecond diffusional rotation of the probe at intermediate stages and in the denatured protein's structure. Also, the large Stokes shift of Nile red allows the changes in the environment of the probe-protein complex in reverse micelles of varying waterpool size to be easily identified in the steady-state fluorescence. This was not seen in earlier work exploiting the intrinsic tryptophan fluorescence of HSA and further demonstrates the complementary information that extrinsic fluorescence probe studies can offer protein science. We discuss the complex acrylamide quenching characteristics of Nile red bound to HSA in terms of the possibility of at least two binding sites for the probe and the effect of acrylamide on the probe-protein structure at very high quencher concentrations.     

Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method

Fluorescence spectrometry based on fluorescence resonance energy transfer (FRET) principle is a simple but effective tool for investigating protein-protein interactions. In this paper, we report a spectrometry to quantify FRET efficiency based on our home-designed spectral probe system and spectral data-processing procedure. In our method, the fluorescence spectrum from each specimen is recorded at two wavelengths 454 and 502 nm. Least-squares linear fitting algorithm is applied directly to decompose the spectra of donor and acceptor under these two wavelengths to obtain FRET efficiency, which takes both spectral intensity and spectral profile into account compared with traditional three-step analysis. This system and the data-processing procedure enabled us to detect the homo-interaction and hetero-interaction of proteins in living cell.