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Spectroscopic approach to capillary-alveolar membrane damage: PATIENTS AND METHODS (3)

Sample preparation and spectroscopic assay: Bronchial washing samples were homogenized for 2 to 3 mins at 100 rpm and centrifuged at 15,000 rpm for 10 mins. Because of the different amounts of fluid recovered, some specimens had to be diluted with distilled water to adjust the final concentration to the optimal spectroscopic range. An aliquot of 3 |il of this bronchial washing was placed on an IR transparent calcium fluoride window, which quickly dried down to form a thin film. This procedure eliminates water, which is a strong IR absorber that dominates the IR spectrum. Spectra were recorded with a FTS-40A (Bio-Rad Laboratories Inc, Cambridge, Massachusetts) IR spectrometer at a resolution of 8 cm-1 over the spectral range of 900 to 4000 cm-1. Data acquisition time was less than 1 min/spectrum, and the procedure can readily be automated to handle batches of samples. Data treatment and statistical analysis: A supervised spectral pattern recognition methodology, linear discriminant analysis, was applied to determine the presence of PENTAS-PAN in the spectrum of the bronchial washing. The methodology had been applied earlier for the classification of synovial fluid spectra to ascertain the nature of the arthritic condition. buy asthma inhaler

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