Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood

This article explores the feasibility of the use of automated microscopy and image analysis to detect the presence of rare fetal nucleated red blood cells (NRBCs) circulating in maternal blood. The rationales for enrichment and for automated image analysis for "rare-event" detection are reviewed. We also describe the application of automated image analysis to 42 maternal blood samples, using a protocol consisting of one-step enrichment followed by immunocytochemical staining for fetal hemoglobin (HbF) and FISH for X- and Y-chromosomal sequences. Automated image analysis consisted of multimode microscopy and subsequent visual evaluation of image memories containing the selected objects. The FISH results were compared with the results of conventional karyotyping of the chorionic villi. By use of manual screening, 43% of the slides were found to be positive (>=1 NRBC), with a mean number of 11 NRBCs (range 1-40). By automated microscopy, 52% were positive, with on average 17 NRBCs (range 1-111). There was a good correlation between both manual and automated screening, but the NRBC yield from automated image analysis was found to be superior to that from manual screening (P=.0443), particularly when the NRBC count was >15. Seven (64%) of 11 XY fetuses were correctly diagnosed by FISH analysis of automatically detected cells, and all discrepancies were restricted to the lower cell-count range. We believe that automated microscopy and image analysis reduce the screening workload, are more sensitive than manual evaluation, and can be used to detect rare HbF-containing NRBCs in maternal blood.     

Simultaneous two-wavelength holographic interferometry in a superorbital expansion tube facility

A new variation of holographic interferometry has been utilized to perform simultaneous two-wavelength measurements, allowing quantitative analysis of the heavy particle and electron densities in a superorbital facility. An air test gas accelerated to 12 km/s was passed over a cylindrical model, simulating reentry conditions encountered by a space vehicle on a superorbital mission. Laser beams with two different wavelengths have been overlapped, passed through the test section, and simultaneously recorded on a single holographic plate. Reconstruction of the hologram generated two separate interferograms at different angles from which the quantitative measurements were made. With this technique, a peak electron concentration of (5.5 +/- 0.5) x 10(23) m(-3) was found behind a bow shock on a cylinder.     

Calculation of I/V characteristics for ion-implanted GaAs MESFETs

The DC characteristics of an ion-implanted MESFET have been calculated from theoretical and measured carrier concentration profiles. The calculation allows the determination of characteristics for planar or recessed gate structures and shows good agreement with experimental devices.     

Near-resonant holographic interferometry and absorption measurements of seeded atomic species in a flame

The development of near-resonant holographic interferometry techniques for use on flows seeded with atomic species is described. A theoretical model for the refractivity that is due to the seed species is outlined, and an approximation to this model is also described that is shown to be valid for practical regimes of interest and allows the number density of the species to be determined without knowledge of line-broadening effects. The details of quantitative number density experiments performed on an air-acetylene flame are given, and a comparison with an alternative absorption-based experiment is made.     

Application of laser-induced breakdown spectroscopy for total carbon quantification in soil samples

The increase of greenhouse gas (i.e., CO(2)) levels in the atmosphere has caused noticeable climate change. Many nations are currently looking into methods of permanent underground storage for CO(2) in an attempt to mitigate this problem. The goal of this work is to develop a process for studying the total carbon content in soils before, during, and after CO(2) injection to ensure that no leakage is occurring or to determine how much is leaking if it is occurring and what effect it will have on the ecosystem between the injection formation and the atmosphere. In this study, we quantitatively determine the total carbon concentration in soil using laser-induced breakdown spectroscopy (LIBS). A soil sample from Starkville, Mississippi, USA was mixed with different amounts of carbon powder, which was used as a calibration for additional carbon in soil. Test samples were prepared by adding different but known amounts of carbon powder to a soil sample and then mixing with polyvinyl alcohol binder before being pressed into pellets. LIBS spectra of the test samples were collected and analyzed to obtain optimized conditions for the measurement of total carbon in soil with LIBS. The total carbon content in the samples was also measured by a carbon analyzer, and the data (average of triplicates) were used as a reference in developing calibration curves for a modified version of the single linear regression model and the multiple linear regression model. The calibration data were then used to determine the total carbon concentration of an unknown sample. This work is intended to be used in the initial development of a miniaturized, field-portable LIBS analyzer for CO(2) leak detection.     

Principal component analysis of TOF-SIMS images of organic monolayers

Principal component analysis (PCA) is a statistical method used to find combinations of variables or factors that describe the most important trends in the data. PCA has been combined with time-of-flight secondary ion mass spectrometry (TOF-SIMS) data to extract new information and find relations between species contained in complex systems. Monolayers of dipalmitoylphosphatidylcholine alone and mixed with palmitoyloleoylphosphatidylglycerol prepared using the Langmuir-Blodgett technique are discussed. PCA software provides image scores and corresponding loadings for each significant principal component. Image plots of the scores show the spatial distribution and intensity of the species defined by the loading plots (mass spectral features). The intensity and resolution of the image scores can result in substantial improvement over that of the regular TOF-SIMS images especially when static conditions are used for small analysis areas. Also, some of the effects of topography and matrix in the images can be removed, allowing for a better presentation of chemical variations.     

Enzyme activity of macrophage migration inhibitory factor toward oxidized catecholamines

Macrophage migration inhibitory factor (MIF) is a relatively small, 12.5-kDa protein that is structurally related to some isomerases and for which multiple immune and catalytic roles have been proposed. MIF is widely expressed in tissues with particularly high levels in neural tissues. Here we show that MIF is able to catalyze the conversion of 3,4-dihydroxyphenylaminechrome and norepinephrinechrome, toxic quinone products of the neurotransmitter catecholamines 3,4-dihydroxyphenylamine and norepinephrine, to indoledihydroxy derivatives that may serve as precursors to neuromelanin. This raises the possibility that MIF participates in a detoxification pathway for catecholamine products and could therefore have a protective role in neural tissues, which as in Parkinson's disease, may be subject to catecholamine-related cell death.