Near-infrared Laser-induced Fluorescence Research Articles

Laser-induced fluorescence spectroscopy on xenon atoms and ions in the magnetic nozzle of a helicon plasma thruster

The dynamics of xenon atoms and ions expanding in the magnetic nozzle (MN) of a helicon plasma thruster is studied by means of near-infrared laser-induced fluorescence spectroscopy on resonant and metastable states. Fluorescence spectra are measured for several operating conditions inside and outside the thruster discharge chamber. In the near-field plume, the relatively intense magnetic field induces Zeeman effect on the probed optical transitions. Hence, modeling of the atomic lineshapes is addressed to accurately compute the Doppler shift and infer the velocity. The first direct measurements of the neutral flow in a MN reveal that atoms are accelerated to supersonic velocities behind the thruster exit. The ions acceleration region extends several centimeters downstream the exit plane. Larger axial ion speeds are attained when the thruster operates at lower mass flow rates and higher levels of input power.

The solvatochromic effects of side chain substitution on the binding interaction of novel tricarbocyanine dyes with human serum albumin

The effects of solvatochromism on protein-ligand interactions have been studied by absorbance and near-infrared laser induced fluorescence (NIR-LIF) spectroscopy. The utility of three novel classes of cyanine dyes designed for this purpose illustrates that the affinity interactions of ligands at the hydrophobic binding pockets of Human Serum Albumin (HSA) are not only dependent on the overall hydrophobic characteristics of the molecules but are highly influenced by the size of the ligands as well. Whereas changes to the chromophore moiety exhibited slight to moderate changes to the hydrophobic nature of these molecules, substitution at the alkyl indolium side chain has enabled us to vary the binding affinity towards serum albumin. Substitution at the indolium side chain among an ethyl to butyl group results in improved binding characteristics and an almost three-fold increase in affinity constant. In addition, replacement of the ethyl side chain with a phenylpropyl group also yielded unique solvotachromic patterns such as increased hydrophobicity and subsequent biocompatibility with the HSA binding regions. Ligand interaction was however inhibited by steric hindrance associated with the bulky phenyl ring system thus affecting the increased binding that could be realized from the improved hydrophobic nature of the molecules. This characteristic change in binding affinity is of potential interest to developing a methodology which reveals information on the hydrophobic character and steric specificity of the binding cavities.

A full analytic potential energy curve for the aΣ+3 state of KLi from a limited vibrational data set

Fourier transform spectra of near-infrared laser-induced fluorescence in (39)K(6)Li show transitions to high vibrational levels of both the X (1)Sigma(+) and a (3)Sigma(+) electronic states. These include 147 transitions into six vibrational levels of the a (3)Sigma(+) state, which lie between 7 and 88 cm(-1) below the dissociation asymptote. Unfortunately, their energies span less than 30% of the well depth. However, fitting those data to eigenvalues of analytical model potential functions whose outer limbs incorporate the theoretically predicted long-range form, V(R) approximately D-C(6)R(6)-C(8)R(8), yields complete, plausible potential curves for this state. The best fits converge to remarkably similar solutions which indicate that D(e)=287(+/-4) cm(-1) and R(e)=4.99(+/-0.09) A for the a (3)Sigma(+) state of KLi, with omega(e)=47.3(+/-1.4) and 44.2(+/-1.5) cm(-1) for (39)K(6)Li and (39)K(7)Li, respectively. Properties of the resulting potential are similar to those of a published ab initio potential and are consistent with those of the analogous states of Li(2), K(2), Na(2), and NaK.

Covalent and noncovalent labeling schemes for near-infrared dyes in capillary electrophoresis protein applications.

Capillary electrophoresis (CE) is experiencing increased use in the field of separation science. Part of its growing popularity of capillary electrophoresis can be attributed to the high efficiency of the separations achievable with the technique, making it an attractive tool for bioanalytical applications. Laser-induced fluorescence (LIF) is a common detection method for CE. One of the problems frequently experienced when using visible LIF detection is matrix autofluorescence which has the effect of degrading the overall sensitivity of the technique. However, the use of near-infrared (NIR) laser induced fluorescence nearly eliminates matrix autofluorescence, as very few molecules have intrinsic fluorescence in this region. This chapter describes the use of covalent and noncovalent labeling schemes for tagging biomolecules with near infrared dyes. To fully appreciate the advantages that the NIR LIF technique can supply, we also review applications that employ detection schemes other than NIR LIF. Specific applications to be discussed include drug-protein studies by CE, as well as capillary electrophoretic immunoassays.

Use of non-covalent labeling in illustrating ligand binding to human serum albumin via affinity capillary electrophoresis with near-infrared laser induced fluorescence detection

This paper demonstrates the use of a near-infrared (NIR) dye as a non-covalent label for human serum albumin (HSA). The dye is a water soluble, heptamethine cyanine dye. The utility of the dye as a tracer illustrating the binding of various drugs to HSA is demonstrated via affinity capillary electrophoresis with near-infrared laser-induced fluorescence detection (ACE-NIR-LIF). Additionally, the factors affecting the separation of relevant species were investigated. The change in quantum yield of the dye upon complexation with HSA was calculated. Spectrophotometric measurements were conducted to study the stoichiometry of the dye albumin complex.

Near-infrared laser-induced fluorescence detection in capillary electrophoresis.

As capillary electrophoresis continues to focus on miniaturization, either through reducing column dimensions or situating entire electrophoresis systems on planar chips, advances in detection become necessary to meet the challenges posed by these electrophoresis platforms. The challenges result from the fact that miniaturization requires smaller load volumes, demanding highly sensitive detection. In addition, many times multiple targets must be analyzed simultaneously (multiplexed applications), further complicating detection. Near-infrared (NIR) fluorescence offers an attractive alternative to visible fluorescence for critical applications in capillary electrophoresis due to the impressive limits of detection that can be generated, in part resulting from the low background levels that are observed in the NIR. Advances in instrumentation and fluorogenic labels appropriate for NIR monitoring have led to a growing number of examples of the use of NIR fluorescence in capillary electrophoresis. In this review, we will cover instrumental components used to construct ultrasensitive NIR fluorescence detectors, including light sources and photon transducers. In addition, we will discuss various types of labeling dyes appropriate for NIR fluorescence and finally, we will present several applications that have used NIR fluorescence in capillary electrophoresis, especially for DNA sequencing and fragment analysis.

Ultrasensitive detection of closely related angiotensin I peptides using capillary electrophoresis with near-infrared laser-induced fluorescence detection.

A novel near-infrared (NIR) fluorescent dye (NN382, LICOR, Inc.) was evaluated as an ultrasensitive peptide-labeling reagent for use with capillary electrophoresis (CE). Six angiotensin I (Ang-I) variants were selected as model peptides for the derivatization and separation studies. The closely related decapeptides were labeled with the NIR dye, separated using CE, and detected by NIR laser-induced fluorescence. Derivatization of the peptides was achieved under aqueous conditions using 2.5-500 pmol of Ang-I in a 50-microL sample (5 x 10(-8)-1 x 10(-5)M), and between 1.3 and 254 amol of the labeled peptides were injected on column. The fluorescence response was linear over a 200-fold range (correlation r > or = 0.9986). The limit of detection (SNR = 3, signal/RMS noise) ranged from 100 to 300 zmol, for the six Ang-I variants. Four of six peptides were resolved from each other and excess dye using capillary zone electrophoresis with a simple 50 mM phosphate run buffer, pH 7.2. Two pairs of coeluting peptides were successfully resolved using micellar electrokinetic chromatography with a nonionic surfactant, Triton X-100. The NIR amine-labeling reagent NN382 is a viable alternative to using visible fluorophores for CE methods requiring high sensitivity.

Interfacing a High-Sensitivity, Near-Infrared Laser-Induced Fluorescence Detector with a Commercial Capillary Electrophoresis Instrument

Interfacing a High-Sensitivity, Near-Infrared Laser-Induced Fluorescence Detector with a Commercial Capillary Electrophoresis Instrument

Ultrasensitive near-infrared laser-induced fluorescence detection in capillary electrophoresis using a diode laser and avalanche photodiode

A sensitive fluorescence detector for capillary electrophoresis consisting of a semiconductor near-infrared diode laser and a single photon avalanche diode (SPAD) is described. The sensitivity of this system was demonstrated by the separation and analysis of four tricarbocyanine dyes using capillary electrophoresis and a running buffer consisting of 98% methanol and 2% water with 40 m M borate (pH 9.4). The LOD for the dye, IR-132, was found to be 4.41 zmol with the dynamic range found to be approximately four orders of magnitude in concentration. Based on the sampling volume of the system, the number of molecules actually detected at this LOD was approximately 27. To further demonstrate the utility of this diode-based detector, various amino acids were derivatized with a highly anionic near-IR labelling dye. The conjugates were separated in a running buffer comprised of predominately methanol and a cationic surfactant added to reverse the electroosmotic flow. The LOD values for various amino acids were found to be in the low zmol range.

Near-infrared laser-induced fluorescence detection in column liquid chromatography. A comparison of various lasers and detection systems II. Pulsed lasers

The applicability of two pulsed lasers, a XeCl-excimer/dye laser and a Nd:YAG/dye laser combination, as excitation sources for near-infrared (NIR) laser-induced fluorescence (LIF) detection in column liquid chromatography (LC) is studied. Using gradient LC, the best detection limit for the model compound, disulphonated aluminum phthalocyanine (AlPcS2), is 4 · 10 −12 M obtained using the excimer/dye laser combination. This value is about 10-fold lower than obtained with the Nd:YAG laser, but 20-fold higher than for CW diode laser excitation, as reported in Part I of this study [ J. Chromatogr. A, 695 (1995) 175]. Because of excitation saturation the optimum results were obtained using only a fraction of the available power. A detailed theoretical discussion is presented which underlines the experimental results and provides insight into the factors that determine the detection limits.

Near-infrared laser-induced fluorescence detection in column liquid chromatography. A comparison of various lasers and detection systems I. Continuous wave lasers

Several lasers, i.e. HeNe lasers, diode lasers and an argon-ion-dye laser combination, are compared as excitation sources for near-infrared laser-induced fluorescence detection in column liquid chromatography. Using a model gradient liquid chromatography system, detection limits for disulphonated aluminum phthalocyanine (A1PcS2) were compared after optimization of the signal processing scheme for each laser. The best detection limit (1.5 · 10 13 M) was obtained for the argon-ion-dye laser combination, applying ratioing of emission and excitation light to reduce flicker noise. Utilizing diode laser excitation there is no need for ratioing so that emission collection can be improved and a detection limit similar to that obtained with teh argon-ion laser can be achieved. As this device is very cheap, the diode laser is the best choice for most applications. If tunability over a large wavelength region is essential, the argon-ion-dye laser combination is a good but expensive alternative.