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PubMed1993

Optical crystallographic characteristics of some USP drugs.

Optical crystallographic data were determined on a variety of newer USP drugs for which data had not been previously obtained. The ingredients in an unknown sample can be identified by polarized light microscopy by mounting small amounts of the material in successive specific refractive index liquids and determining refractive indices, optic sign, type of extinction, estimated 2V angle, and other optical characteristics for each ingredient present. This information can then be compared with the data listed on known drugs and excipients and all of the ingredients can be identified. To aid in the identification of unknown drugs, optical crystallographic data obtained on 52 of the newer USP drugs are presented.

Crystallography↗
PubMed2005

Measurement of the coagulation dynamics of bovine liver using the modified microscopic Beer-Lambert law.

BACKGROUND AND OBJECTIVES: During heating, the optical properties of biological tissues change with the coagulation state. In this study, we propose a technique, which uses these changes to monitor the coagulation process during laser-induced interstitial thermotherapy (LITT). STUDY DESIGN/MATERIALS AND METHODS: Untreated and coagulated (water bath, temperatures between 35 degrees C and 90 degrees C for 20 minutes.) samples of bovine liver tissue were examined using a Nd:YAG (lambda = 1064 nm) frequency-domain reflectance spectrometer. We determined the time integrated intensities (I(DC)) and the phase shifts (Phi) of the photon density waves after migration through the tissue. From these measured quantities, the time of flight (TOF) of the photons and the absorption coefficients of the samples were derived using the modified microscopic Beer-Lambert law. RESULTS: The absorption coefficients of the liver samples decreased significantly with the temperature in the range between 50 degrees C and 70 degrees C. At the same time, the TOF of the investigated photos was found increased indicating an increased scattering. The coagulation dynamics could be well described using the Arrhenius formalism with the activation energy of 106 kJ/mol and the frequency factor of 1.59 x 10(13)/second. CONCLUSIONS: Frequency-domain reflectance spectroscopy in combination with the modified microscopic Beer-Lambert (MBL) is suitable to measure heat induced changes in the absorption and scattering properties of bovine liver in vitro. The technique may be used to monitor the coagulation dynamics during local thermo-coagulation in vivo.

Algorithms↗
PubMed2006

In vivo quantitative three-dimensional motion mapping of the murine myocardium with PC-MRI at 17.6 T.

This work presents a method that allows for the assessment of 3D murine myocardial motion in vivo at microscopic resolution. Phase-contrast (PC) magnetic resonance imaging (MRI) at 17.6 T was applied to map myocardial motion in healthy mice along three gradient directions. High-resolution velocity maps were acquired at three different levels in the murine myocardium with an in-plane resolution of 98 mum, a slice thickness of 0.6 mm, and a temporal resolution of 6 ms. The applied PC-MRI method was validated with phantom experiments that confirmed the correctness of the method with deviations of <1.7%. Myocardial in-plane velocities between 0.5 cm/s and 2.2 cm/s were determined for the healthy murine myocardium. Through-plane velocities of 0.1-0.83 cm/s were measured. Velocity data was also used to calculate the myocardial twist angle during systole at different slices in the short-axis view.

Algorithms↗
PubMedDate not supplied

Carbohydrate analysis of floral nectar using medium infrared.

An instrumental method based on a chemometric model of the medium region of the infrared (MIR) was developed to analyse total sugar content and the proportions of glucose, fructose and sucrose. In order to construct the model, a set of 127 standard aqueous solutions of different sugars in the concentration range 0-20% (w/v) were prepared and analysed in the interval 4900-700 cm(-1). The MIR was transformed by normalisation, correction of baseline using the second derivative, and suppression of the signals of water and carbon dioxide. The region between 1150 and 950 cm(-1) showed the highest correlation between signal and concentration. The correlation coefficient for total sugar content was 0.956, whilst those for glucose, fructose and sucrose were 0.982, 0.972 and 0.992, respectively. The method was validated using a set of 28 samples of nectar which had been assayed by chromatographic and refractometric methods. The method shows potential utility for the prediction of nectar sugar components.

Cactaceae↗
PubMed2004

Ultrafast dynamics of a solution in spatially restricted environments studied by photothermal spectroscopies.

The ultrafast dynamics of a solution in spatially restricted environments was studied by using the ultrafast transient lens (UTL) method. The UTL method is used to monitor the molecular dynamics of a solution by means of a change in the refractive index, which is advantageous for investigating the molecular dynamics of restricted systems. We investigated the photoisomerization of azobenzene derivatives in cyclodextrin nanocavities and revealed how the confinement affects the photoisomerization dynamics and yields. We also studied the relaxation dynamics of photo-excited auramine O (AuO) in a water/aerosol-OT/n-heptane reversed micelle. Both the perturbed properties of the included water and the interactions between AuO and the interface of the reversed micelle strongly appeared to affect the relaxation dynamics. At the same time, we observed a change in the refractive index suggesting a structural change of the micelles in the picosecond region that could not be detected by transient absorption spectroscopy. In addition, we developed the total internal reflection UTL (TIR-UTL) method to monitor the ultrafast molecular dynamics at the liquid interface. The relaxation dynamics of photoexcited AuO at the silica/water interface were observed with subpicosecond time resolution, and it was revealed that the interaction with the interface strongly inhibited the relaxation process. These results demonstrated the advantages of the UTL method for investigating the molecular dynamics of a solution in spatially restricted environments.

Azo Compounds↗
PubMed2006

Photorefractive effect of ferroelectric liquid crystals.

This paper reviews our recent work on the photorefractive effect of ferroelectric liquid crystals (FLCs). The photorefractive effect is defined as the optical modulation of the refractive index of a medium as a result of a variety of processes. The interference of two laser beams in a photorefractive material establishes a refractive index grating. This phenomenon enables the creation of different types of photonic applications. FLCs exhibit fast electric field response, and the orientation of the molecular axis of FLCs changes its direction according to the change in direction of the spontaneous polarization (Ps). When two laser beams interfere in a photoconductive FLC, an orientational grating is formed. The mechanism of the formation of the grating is based on the response of the Ps to the photoinduced internal electric field. The time of formation of the refractive index grating is significantly shorter in FLC materials.

Lasers↗
PubMed1999

Application of a thermodynamic model to the prediction of phase separations in freeze-concentrated formulations for protein lyophilization.

Many of the compounds considered for use in pharmaceutical formulations demonstrate incompatibilities with other components at high enough concentrations, including pairs of polymers, polymers and salts, or even proteins in combination with polymers, salts, or other proteins. Freeze concentration can force solutions into a region where incompatibilities between solutes will manifest as the formation of multiple phases. Such phase separation complicates questions of the stability of the formulation as well as labile components, such as proteins. Yet, phase separation events are difficult to identify by common formulation screening methods. In this report, we use the osmotic virial expansion model of Edmond and Ogston (1) to describe phase-separating behavior of ternary aqueous polymer solutions. Second osmotic virial coefficients of polyethylene glycol 3350 (PEG) and dextran T500 were measured by light scattering. Assuming an equilibrium between ice and water in the freeze-concentrated solution, a degree of freeze concentration can be estimated, which, when combined with the phase separation spinodal, describes a "phase separation envelope" in which phase separation tendencies can be expected in the frozen solution. The phase separation envelope is bounded at low temperatures by the glass transition temperature of the freeze-concentrated solution. Scanning electron microscopic images and infrared spectroscopy of protein structure are provided as experimental evidence of the phase separation envelope in a freeze-dried system of PEG, dextran, and hemoglobin.

Dextrans↗
PubMed1995

Determination of sulfamethazine residues in milk by a surface plasmon resonance-based biosensor assay.

The use of antibiotics and chemotherapeutics in animal husbandry has led to the occurrence of veterinary drug residues in all types of food of animal origin. Due to the specification of toxicologically based maximum residue levels for a large number of substances, existing control strategies need even faster and more sensitive methods to meet new and more rigorous regulations. The applicability of an immunosensor device for biospecific interaction analysis was investigated and the development of an assay for analysis of sulfamethazine (SMZ) in milk is described. SMZ was covalently immobilized to a carboxymethyldextran-modified gold film. Spiked samples with known concentrations of SMZ were prepared in HBS buffer and skim and raw milk for construction of standard curves. Polyclonal antibodies against SMZ were added to the sample and the immobilized surface was used to determine the amount of free antibodies by surface plasmon resonance detection. After each measurement the surface was regenerated by NaOH and HCl. In milk, the mean relative standard deviation of the assay was approximately 2% and the limit of detection less than 1 ppb. By introduction of a secondary sheep anti-rabbit antibody, the use of specific antibody could be reduced. Milk samples from the individual cow, herd, and tanker levels were analyzed and the relative standard deviations within each sample category were 4.4, 2.4, and 2.2%, respectively. The effect of some potential interferences, e.g., high somatic cells, bacterial contamination, and preservatives, was investigated. The results were not influenced in such a way that the risk for so-called false-positive findings was obvious.

Animals↗
PubMed1995

Effects of solutes on optical properties of biological materials: models, cells, and tissues.

Perturbations of scattering background for absorbance measurements by photon diffusion techniques complicate algorithms used for determining the concentration of blood and the saturation of hemoglobin in tissues. In order to better define these perturbations, we have undertaken a study of the effect of solutes, some of physiological importance, upon the scattering of three types of model systems: a lipid vessel suspension (Intralipid), a cell suspension (Baker's yeast), and a tissue (perfused liver). A simple formula relates absorbancy change to proportional changes of the input/output separation rho and the square root of mu a and mu's in relation to a relevant model system. Thus, absorbance changes at 850 nm slopes and intercepts are measured as a function of rho; the absorption and reduced scattering coefficient, mu a and mu's, are calculated. We studied each of these cases as a function of the perturbation induced by low-molecular-weight polyhydroxy solutes, generally sugars (mannitol, fructose, sucrose, and glucose), alcohols (propanediol and methanol), and electrolytes (sodium and potassium chloride). Dilatometric studies indicate volume changes of the solvent system and afford correction factors. The slopes are approximately +/- 0.5 x 10(-4) OD/mM solute per centimeter separation of input/output per percent scatter (yeast or Intralipid) and indicate possible physiological detection of solutes in tissues in the millimolar range. The large optical effects of temperature upon the solute effect on model systems and of osmotic and perfusion pressures on the perfused liver further complicate the possibility of quantitative in vivo studies of these solutes.

Animals↗
PubMed1995

Determination of local refractive index for protein and virus crystals in solution by Mach-Zehnder interferometry.

To establish the importance of, and quantitatively evaluate, the macromolecular concentration gradients in the neighborhood of growing protein, virus, and nucleic acid crystals, a convenient, accurate, and nonintrusive method has been devised. This approach should prove particularly relevant in the rigorous comparison of crystals grown in a conventional laboratory setting with those grown in a microgravity environment. The method is based on precise determination of the local refractive index using Mach-Zehnder interferometry. Presented here are data for five protein and three virus systems. From data for these and other systems, optical monitoring experiments to measure local growth conditions and growth kinetics in liquid-liquid diffusion, batch, and vapor diffusion crystal growth experiments can be designed.

Interferometry↗
PubMed1998

Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications.

Submicroscopic gold particle suspensions scatter colored light when illuminated with white light, and we have observed that a light-scattering gold particle suspension has the same appearance as a fluorescing solution. Thus, when illuminated by a narrow beam of white light, a 40-nm gold sol displays a clear (not cloudy), green scattered light (Tyndall) beam and has the same appearance as a fluorescing fluorescein solution. These, as well as other, observations have suggested to us that, in general, light-scattering particles can be treated as fluorescent analogs and used as fluorescent analog tracers in immuno- and DNA probe assays as well as in cell and molecular biology studies. Light-scattering particles are advantageous in these applications because particles such as gold and silver have very high light-scattering powers, which allows these particles to be easily detected, by light-scattering, at particle concentrations as low as 10(-16) M. The scattered light can be detected by the unaided eye for qualitative measurements or with a simple light-sensitive detector for quantitative measurements. Moreover, individual particles can be easily detected by eye or a video camera using a simple light microscope with a proper illuminating system. In addition, submicroscopic particles which scatter blue, green, yellow, orange, or red light can be readily synthesized. Antibodies, DNA probes, and other tracer substances can be readily attached to gold and other particles without altering their light-scattering properties. In this article we present the theory which allows one to predict the light-scattering properties of particles of different sizes and compositions and identify those particle sizes and compositions which appear most adequate for particular applications. Furthermore, we calculate molar extinction coefficients and emission efficiencies for particles of different sizes and compositions which allows us to compare the light-producing powers of these particles with those of well known fluorescent tracers. A 60-nm gold particle, for example, is equivalent to about 3 x 10(5) fluorescein molecules. Very simple, easy to use, low-cost, ultrasensitive immuno- and DNA probe assays can be developed using light-scattering particles as fluorescent analog tracers. Single particles can be detected on cell surfaces and inside cells using light microscopy techniques with proper illumination as described in the article. At high particle densities, particle-labeled cells have the same appearance as fluorescent cells.

Fluorescence↗
PubMed1998

Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications.

In a companion article we present the idea that submicroscopic light-scattering particles, such as gold and silver particles, can be used as fluorescent analog tracers in biological and clinical applications. The light-emitting power and scattered light color of the particles can be adjusted by changing particle composition or diameter. Using Rayleigh and Mie light-scattering theory, we calculated the absorbance and light-scattering properties and power of particles of different diameters and selected compositions and compared them to the properties of fluorophores. In the present article, we evaluate experimentally the optical properties of particles of selected compositions and sizes and compare the results with the theoretically calculated values. We also discuss the methods which we use to measure the light-scattering properties of particles in suspension and to view individual particles by light microscopy. We also outline examples which demonstrate the use of light-scattering particles as fluorescent analogs in biological and clinical applications.

Calibration↗
PubMed2000

Determination of the refractive index increments of small molecules for correction of surface plasmon resonance data.

The refractive index increments (RIIs) of several important low-molecular-weight compounds that bind to DNA or RNA were determined with a differential refractometer for correction of data obtained on surface plasmon resonance (SPR) biosensors. Although the ability to investigate small molecule-macromolecule interactions by SPR is relatively new, the technique is rapidly becoming a primary method to screen focused combinatorial libraries and to quantitatively characterize the interactions between compounds identified as binders and target macromolecules. The most widely used SPR analysis software, BIAevaluation (Biacore, Inc.), assumes that the RIIs of ligand and macromolecule are identical. While the assumption is reasonable for studies involving like molecules such as protein-protein interactions, results presented here demonstrate that RII values for small molecules can be significantly different than those of protein or nucleic acid receptors. The results also show that the RII values can vary greatly depending on the structure of the small molecule. Indeed, the RIIs of the molecules investigated here differ by a factor of 2. Any difference in the RII of interacting molecules must be considered for complete analysis of SPR data. Failure to correct for RII differences can result in serious error in data interpretation, especially for systems involving a ligand:receptor stoichiometry greater than 1. The results serve as the beginning of an SPR correction database for the RIIs of small molecules. Additionally, the results can be used to approximate the RIIs of a variety of other small molecules.

Molecular Weight↗
PubMed2001

Determination of carbohydrate contents from excess light scattering.

In the light scattering technique, glycosylation gives rise to excess light scattering for glycoproteins. Assuming additivity of refractive index and using an appropriate refractive index increment for carbohydrate, one can determine the degree of glycosylation from the excess light scattering. Here we have used size-exclusion chromatography in combination with online light scattering, UV absorbance, and refractive index. The results show that the technique accurately determines the carbohydrate content of recombinant stem cell factor.

Animals↗
PubMed1994

Colloid osmotic pressure of steer alpha- and beta-crystallins: possible functional roles for lens crystallin distribution and structural diversity.

This study addresses the general mechanisms whereby the major cytoplasmic proteins from the adult bovine lens contribute both to transparency and maintenance of the refractive index gradient across the lens. Colloid osmotic properties and quaternary structure were measured for alpha- and beta-crystallins isolated from the steer lens, including low-molecular-weight crystallins from the cortex (alpha Le and beta L) and nucleus (alpha Ln) and high-molecular-weight crystallins from the nucleus (alpha H and beta H). In electron microscopic images of rotary-shadowed preparations alpha Le appears as spherical particles 16 nm in diameter, alpha Ln appeared as individual spheres or small aggregates of spherical subunits, alpha H contained large irregular aggregates as large as 180 nm, and both beta L and beta H appeared as elliptical particles of 7-9 nm diameter. Secondary osmometry showed that for all these crystallins colloid osmotic pressure increased monotonically in a non-linear fashion with protein concentration. For the alpha-crystallins, osmotic pressure rose more steeply with concentration for alpha Le than for either alpha Ln or alpha H, so that at 0.3 g ml-1 at 0.1 M ionic strength, the colloid osmotic pressure of alpha Le, alpha Ln and alpha H were approximately 2.6 x 10(5) dyn cm-2, 1.6 x 10(5) dyn cm-2 and 1.0 x 10(5) dyn cm-2, respectively. In a similar manner, osmotic pressure rose more steeply with concentration of beta L than for beta H, so that at 0.3 g ml-1 at 0.1 M ionic strength the colloid osmotic pressures of beta L and beta H were 2.6 x 10(5) dyn cm-2 and 1.1 x 10(5) dyn cm-2, respectively. The osmotic pressure of alpha Le dropped as ionic strength was increased from 0.02 to 0.4 M. For beta L and beta H, osmotic pressure dropped as ionic strength was increased from 0.02 to 0.1 M but was nearly the same at 0.1 M and 0.4 M ionic strength. The data for steer alpha Ln and beta H were similar to previous reports for calf cortical alpha L and beta-crystallins, respectively. The osmotic pressure isotherms for alpha Le, beta L and that previously reported for steer cortical extract were nearly identical, whereas the nuclear crystallins (alpha Ln, alpha H or beta H) generated slightly higher pressures than those previously reported for steer nuclear crystallin extracts. In all cases, osmotic pressure rose more steeply with concentration for the cortical crystallins than for the nuclear crystallins.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1998

Investigation of dispersion effects in ocular media by multiple wavelength partial coherence interferometry.

We report on quantitative measurements of group refractive indices and group dispersion in water and in human ocular media such as the cornea, the aqueous humor, the lens, artificial intraocular lenses, as well as a total value averaged over the media along the axial eye length of normal subjects and pseudophakic patients in vivo using dual beam partial coherence interferometry. Different optical thickness values due to the dispersion of the cornea are demonstrated using two spectrally displaced light sources. The displacement can be used to indirectly calculate the group dispersion of the human cornea in the spectral region between 810 nm and 860 nm. If the object under investigation is dispersive, resolution is limited due to a broadening of the detected signals. This broadening increases with group dispersion, i.e., the extent to which the group refractive index of the medium varies with wavelength and thickness of the tissue under investigation as well as with the spectral bandwidth of the light source. Measurements of the group dispersion in the cornea, lens and vitreous of pseudophakic and normal human eyes, show that the cornea and the lens are more dispersive than water-by a factor of about 5 and 2, respectively-in the investigated spectral region. The cornea is approximately threefold more dispersive than the human crystalline lens, the aqueous humor is less dispersive than water and the group dispersion of all ocular components together, averaged over the axial length of normal and pseudophakic eyes, was only slightly higher compared to that of water. Since the highly dispersive cornea and lens together have only a thickness of about one sixth of that of the axial eye length, it seems that their contribution to the group dispersive effect along the whole axial eye length is only small.

Aqueous Humor↗