RESUMEN
The standard practice in blood banks worldwide involves storage of red blood cells (RBCs) in plastic bags until they are needed for transfusion. During storage, the cells gradually degrade in functionality, a condition described as RBC storage lesion. Standard analytical techniques cannot assess the blood quality without breaching the sterility of the transfusion bag. In this study, we employed a commercially available spatially offset Raman spectroscopy (SORS) system using a custom designed protocol to non-invasively explore the biochemical changes in RBC concentrate of healthy donors over a storage period of approximately 42 days in standard transfusion bags, under standard storage conditions. The results reveal an increase in the oxygenation state of haemoglobin over the storage period for all donors, but different profiles for each donor. This study demonstrates the feasibility of acquiring consistent biochemical information relevant to the quality of stored blood, in situ through sealed blood transfusion bags using a commercially available instrument.
Asunto(s)
Conservación de la Sangre/efectos adversos , Transfusión Sanguínea/instrumentación , Eritrocitos/metabolismo , Hemoglobinas/metabolismo , Humanos , Masculino , Oxígeno/sangre , Espectrometría Raman/métodosRESUMEN
After being separated from (donated) whole blood, red blood cells are suspended in specially formulated additive solutions and stored (at 4 °C) in polyvinyl chloride (PVC) blood-bags until they are needed for transfusion. With time, the prepared red cell concentrate (RCC) is known to undergo biochemical changes that lower effectiveness of the transfusion, and thus regulations are in place that limit the storage period to 42 days. At present, RCC is not subjected to analytical testing prior to transfusion. In this study, we use Spatially Offset Raman Spectroscopy (SORS) to probe, non-invasively, the biochemistry of RCC inside sealed blood-bags. The retrieved spectra compare well with conventional Raman spectra (of sampled aliquots) and are dominated by features associated with hemoglobin. In addition to the analytical demonstration that SORS can be used to retrieve RCC spectra from standard clinical blood-bags without breaking the sterility of the system, the data reveal interesting detail about the oxygenation-state of the stored cells themselves, namely that some blood-bags unexpectedly contain measurable amounts of deoxygenated hemoglobin after weeks of storage. The demonstration that chemical information can be obtained non-invasively using spectroscopy will enable new studies of RCC degeneration, and points the way to a Raman-based instrument for quality-control in a blood-bank or hospital setting.
Asunto(s)
Transfusión Sanguínea , Eritrocitos/química , Cloruro de Polivinilo , Manejo de Especímenes , Espectrometría Raman , Hospitales , Humanos , Embalaje de ProductosRESUMEN
We propose to employ the technique of femtosecond pulse shaping for improving the performance of the recently suggested method of complete characterization of molecular vibrations, in which both the amplitude and phase of the laser induced vibrational coherence are detected with high resolution. The amplitude-phase information is retrieved from the cross-correlation frequency resolved optical gating of Raman modes. By creating rich interference pattern in the measured two-dimensional spectrogram of coherent anti-Stokes Raman scattering we enhance the accuracy of the retrieved spectral and temporal response and increase the robustness of the method against noise.
RESUMEN
This study presents a new method of image signal-to-noise ratio (SNR) enhancement by utilizing a newly developed 2D two-point maximum entropy regularization method (TPMEM). When utilized as an image filter, it is shown that 2D TPMEM offers unsurpassed flexibility in its ability to balance the complementary requirements of image smoothness and fidelity. The technique is evaluated for use in the enhancement of x-ray computed tomography (CT) images of irradiated polymer gels used in radiation dosimetry. We utilize a range of statistical parameters (e.g. root-mean square error, correlation coefficient, error histograms, Fourier data) to characterize the performance of TPMEM applied to a series of synthetic images of varying initial SNR. These images are designed to mimic a range of dose intensity patterns that would occur in x-ray CT polymer gel radiation dosimetry. Analysis is extended to a CT image of a polymer gel dosimeter irradiated with a stereotactic radiation therapy dose distribution. Results indicate that TPMEM performs strikingly well on radiation dosimetry data, significantly enhancing the SNR of noise-corrupted images (SNR enhancement factors >15 are possible) while minimally distorting the original image detail (as shown by the error histograms and Fourier data). It is also noted that application of this new TPMEM filter is not restricted exclusively to x-ray CT polymer gel dosimetry image data but can in future be extended to a wide range of radiation dosimetry data.
Asunto(s)
Algoritmos , Geles/efectos de la radiación , Modelos Químicos , Polímeros/efectos de la radiación , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Radiometría/métodos , Tomografía Computarizada por Rayos X/métodos , Simulación por Computador , Entropía , Geles/química , Modelos Estadísticos , Polímeros/química , Dosis de Radiación , Radiometría/instrumentación , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Tomografía Computarizada por Rayos X/instrumentaciónRESUMEN
The development and characterization of a new instrument for solid sampling which couples IR laser desorption followed by UV laser photo-ionization and analysis using an ion trap mass spectrometer has been investigated. For calibration, a new type of solid sample preparation involving activated charcoal as the solid substrate was used. This solid sample provided a steady signal for several thousand laser shots, which allowed optimization of the experimental procedure. It was found that both the IR and UV intensity and the delay between them play an important role in both the magnitude and type of signals observed. A method of gas phase accumulation with multiple laser shots was examined. Finally, this technique was demonstrated to be effective in providing direct qualitative information for N.I.S.T. SRM 1944 river sediment sample with no sample pre-treatment.
Asunto(s)
Hidrocarburos Policíclicos Aromáticos/análisis , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/instrumentación , Contaminantes Químicos del Agua/análisis , Carbón Orgánico/análisis , Rayos Infrarrojos , Rayos Láser , Pirenos/análisis , Reproducibilidad de los Resultados , Programas Informáticos , Rayos UltravioletaRESUMEN
Many techniques have been developed to investigate the chemistry associated with brain activity. These techniques generally fall into two categories: fast techniques with species-limited sensitivity; and generally slower techniques with broader species sensitivity. Therefore, a need exists for a fast, minimally invasive technique that is sensitive to a wide array of biologically relevant compounds in order to measure chemical brain events in real time. The work presented here describes the development of a novel spectroscopic neurotransmitter probe for the rapid and simultaneous detection of a variety of neurotransmitters. A fiber-optic-linked Raman and tunable ultraviolet resonance Raman system was assembled with custom designed optical fiber probes. Using this system, the ultraviolet resonance Raman spectra of some small-molecule and peptide neurotransmitters were measured in-vitro with a fiber-optic probe and are reported here for the first time. The probe has furthermore been used to measure neurotransmitter secretions obtained from depolarized rat pheochromocytoma (PC12) cells. These results demonstrate the general utility of this approach which, due to the fiber-optic implementation, could potentially also be applied to in-vivo neurotransmitter determinations.
Asunto(s)
Acetilcolina/análisis , Monoaminas Biogénicas/análisis , Neuropéptidos/análisis , Espectrometría Raman/métodos , Animales , Tecnología de Fibra Óptica/instrumentación , Fibras Ópticas , Células PC12 , Ratas , Espectrofotometría Ultravioleta/métodos , Espectrometría Raman/instrumentaciónRESUMEN
Identification of individual components in biological mixtures can be a difficult problem regardless of the analytical method employed. In this work, Raman spectroscopy was chosen as a prototype analytical method due to its inherent versatility and applicability to aqueous media, making it useful for the study of biological samples. Artificial neural networks (ANNs) and the classical least-squares (CLS) method were used to identify and quantify the Raman spectra of the small-molecule neurotransmitters and mixtures of such molecules. The transfer functions used by a network, as well as the architecture of a network, played an important role in the ability of the network to identify the Raman spectra of individual neurotransmitters and the Raman spectra of neurotransmitter mixtures. Specifically, networks using sigmoid and hyperbolic tangent transfer functions generalized better from the mixtures in the training data set to those in the testing data sets than networks using sine functions. Networks with connections that permit the local processing of inputs generally performed better than other networks on all the testing data sets. and better than the CLS method of curve fitting, on novel spectra of some neurotransmitters. The CLS method was found to perform well on noisy, shifted, and difference spectra.
Asunto(s)
Análisis de los Mínimos Cuadrados , Redes Neurales de la Computación , Neurotransmisores/análisis , Acetilcolina/análisis , Acetilcolina/líquido cefalorraquídeo , Animales , Masculino , Ratas , Espectrometría RamanRESUMEN
A technique is presented to simply and effectively decompose the perturbation domain in two-dimensional (2D) correlation maps calculated on a given set of vibrational spectra. Decomposition of the perturbation domain exposes a wealth of kinetic information complementary to the information extracted from conventional 2D correlation spectroscopy. It is shown that the technique produces "perturbation profile maps" that can be utilized in both the interpretation of the conventional 2D correlation maps and the independent kinetic analysis of the given system. Discrimination between spectral features exhibiting similar, but not identical, dynamics is facilitated by the decomposition, and spectral features exhibiting identical dynamics over the perturbation interval are quickly identified. Spectral features exhibiting similar dynamics over only a sub-range of the full perturbation are also identifiable. Interpretation of phase information illuminated in synchronous and asynchronous maps is simplified. Comparison between similar spectral features present in different samples is facilitated with the technique. The simplicity and ease of implementation of the technique make decomposition of the perturbation domain a valuable addition to the tools available in 2D correlation analysis.
RESUMEN
Generalized two-dimensional correlation spectroscopy offers great scope for revealing the behavior of relationships between components of a system under empirical study. We have developed methods that aid in the interpretation of two-dimensional correlation spectroscopy. These methods include reference patterns for two-dimensional correlation and correlation coefficient maps, their superposition and joint interpretation, and the use of delta functions to decompose them in the perturbation domain. We show how their joint use permits discrimination between similar two-dimensional correlation map features on the basis of different correlation coefficients. We also show how the decomposition of maps into the perturbation domain reflects the dynamic behavior of spectral features over the course of the perturbation and permits discrimination between otherwise highly similar two-dimensional correlation cross-peaks. These approaches simplify the interpretation of two-dimensional correlation spectroscopy maps and facilitate access to their rich information content.
RESUMEN
Vibrational spectra often require baseline removal before further data analysis can be performed. Manual (i.e., user) baseline determination and removal is a common technique used to perform this operation. Currently, little data exists that details the accuracy and precision that can be expected with manual baseline removal techniques. This study addresses this current lack of data. One hundred spectra of varying signal-to-noise ratio (SNR), signal-to-baseline ratio (SBR), baseline slope, and spectral congestion were constructed and baselines were subtracted by 16 volunteers who were categorized as being either experienced or inexperienced in baseline determination. In total, 285 baseline determinations were performed. The general level of accuracy and precision that can be expected for manually determined baselines from spectra of varying SNR, SBR, baseline slope, and spectral congestion is established. Furthermore, the effects of user experience on the accuracy and precision of baseline determination is estimated. The interactions between the above factors in affecting the accuracy and precision of baseline determination is highlighted. Where possible, the functional relationships between accuracy, precision, and the given spectral characteristic are detailed. The results provide users of manual baseline determination useful guidelines in establishing limits of accuracy and precision when performing manual baseline determination, as well as highlighting conditions that confound the accuracy and precision of manual baseline determination.
Asunto(s)
Algoritmos , Interpretación Estadística de Datos , Modelos Químicos , Modelos Estadísticos , Análisis Espectral/métodos , Simulación por Computador , Variaciones Dependientes del Observador , Control de Calidad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Análisis Espectral/normas , Procesos EstocásticosRESUMEN
Two types of wide dynamic range integrating read-out systems for photomultiplier tubes have been developed. One system is based on a voltage-to-frequency converter-counter system and the second is based on summing a large number of analogue-to-digital converter values. Both systems operate under microprocessor control. The microprocessor controls and sequences the acquisition of replicate integrated measurements and software is provided for calculation of background-corrected values, standard deviations for up to 32 replicate values and signal-to-noise ratios. Development and implementation of this software capability in a small system is greatly facilitated by use of a hardware floating-point package. All program software is stored in erasable programmable read-only memory. This system has been particularly useful for the acquisition of data from a single-channel inductively-coupled plasma atomic-emission spectrometer.
RESUMEN
The best analytical line for determination of a specified analyte is selected from a set of lines on the basis of the least interference in a particular sample matrix, by several cycles of mathematical analysis. Unsuitable lines are rejected in the first few cycles, the best lines being retained until last. A multivariate analysis after each cycle provides an updated estimate of the analyte concentration. This process is performed without reference to spectral tables.
RESUMEN
Some microcomputers have been evaluated in terms of their features and capabilities for basic laboratory data-acquisition tasks. The systems studied include the SDK-80 (8080), SDK-85 (8085), and KIM-1 (6502) single-board microcomputers, and the PET consumer/hobby-market microcomputer. Data-acquisition software benchmarks were tested on all the systems, and inexpensive analogue-to-digital and digital-to-analogue hardware subsystems were developed for the PET. The acquisition rates for 10-bit data under machine-language control range from 15 to 25 kHz, but under high-level (BASIC) language-control drop to about 100 Hz.
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A method is described to determine selenium in biological material, based on cathodic stripping voltammetry. Following wet ashing, the selenium was extracted into benzene as the 3',4'-diaminophenylpiazselenol. The selenium was subsequently back-extracted into dilute acid for analysis. Analyses of NBS Bovine Liver demonstrated that the method was capable of recovering 96+/-9% of the selenium present. The detection limit and working range were 3 ng/g and 0-10,000 ng/g, respectively. The method was also applied to the determination of selenium in rapeseed oils and seed.
Asunto(s)
Hígado/análisis , Selenio/análisis , Animales , Bovinos , Electroquímica , Métodos , Microquímica , Aceites/análisis , Semillas/análisisRESUMEN
We investigated the performance of fiber-optic resonance Raman probes with a series of experiments to determine the working curves of such probes using model analytes and to investigate the effects of absorbing media. A computer model designed to simulate these experiments is presented, and numerical results are found to be in agreement with the experimental data. Design considerations resulting from these studies are discussed, and novel designs for overcoming problems of coupling efficiency, damage threshold, and sensitivity in absorbing samples are presented. These findings are applied to the design of fiber-optic probes for ultraviolet resonance Raman spectroscopy involving nanosecond pulsed-ultraviolet excitation (225 and 266 nm). These probes have been used to collect what is, to our knowledge, the first reported fiber-optic-linked ultraviolet resonance Raman spectra of tryptophan and DNA.
RESUMEN
The ability of ultraviolet resonance Raman spectroscopy (UVRRS) to determine structural, environmental, and analytical information concerning low-concentration aqueous biomolecules makes it a powerful bioanalytical and biophysical technique. Unfortunately, its utility has been limited by experimental requirements that preclude in situ or in vivo studies in most cases. We have developed the first high-performance fiber-optic probes suitable for long-term use in pulsed UVRRS applications in the deep- UV (DUV, 205-250 nm). The probes incorporate recently developed improved ultraviolet (IUV) fibers that do not exhibit the rapid solarization and throughput decay that previously hampered the use of optical fibers for delivering pulsed, DUV light. A novel 90 degrees mirrored collection geometry is used to overcome the inner-filtering effects that plague flush-probe geometries. The IUV fibers are characterized with respect to their efficacy at transmitting pulsed, DUV laser light, and prototype probes are used to obtain pulsed UVRRS data of aromatic amino acids, proteins, and hormones at low concentrations with 205-240-nm pulsed excitation. Efficient probe geometries and fabrication methods are presented. The performance of the probes in examining resonance-enhanced Raman signals from absorbing chromophores is investigated, and the optimal excitation wavelength is shown to be significantly red-shifted from the maximum of the resonance Raman enhancement profile. Generally applicable procedures for determining optimal experimental conditions are also introduced.