Infrared and NIR raman spectroscopy in medical microbiology

FT-IR and FT-NIR Raman spectra of intact microbial cells are highly specific, fingerprint-like signatures which can be used to (i) discriminate between diverse microbial species and strains, (ii) detect in situ intracellular components or structures such as inclusion bodies, storage materials or endospores, (iii) detect and quantify metabolically released CO2 in response to various different substrates, and (iv) characterize growth-dependent phenomena and cell-drug interactions. The characteristic information is extracted from the spectral contours by applying resolution enhancement techniques, difference spectroscopy, and pattern recognition methods such as factor-, cluster-, linear discriminant analysis, and artificial neural networks. Particularly interesting applications arise by means of a light microscope coupled to the spectrometer. FT-IR spectra of micro-colonies containing less than 10(3) cells can be obtained from colony replica by a stamping technique that transfers micro-colonies growing on culture plates to a special IR-sample holder. Using a computer controlled x,y-stage together with mapping and video techniques, the fundamental tasks of microbiological analysis, namely detection, enumeration, and differentiation of micro-organisms can be integrated in one single apparatus. FT-IR and NIR-FT-Raman spectroscopy can also be used in tandem to characterize medically important microorganisms. Currently novel methodologies are tested to take advantage of the complementary information of infrared and Raman spectra. Representative examples on medically important microorganisms will be given that highlight the new possibilities of vibrational spectroscopies.