Martin, Frank and Kerns, Jemma and Llabjani, Valon and Martin-Hirsch, Pierre L. and Patel, Imran I. and Trevisan, Julio and Fullwood, Nigel J. and Walsh, Michael J. (2010) Distinguishing cell types or populations based on the computational analysis of their infrared spectra. Nature Protocols, 5 (11). pp. 1748-1760. ISSN 1754-2189
Full text not available from this repository.Abstract
Infrared (IR) spectroscopy of intact cells results in a fingerprint of their biochemistry in the form of an IR spectrum; this has given rise to the new field of biospectroscopy. This protocol describes sample preparation (a tissue section or cytology specimen), the application of IR spectroscopy tools, and computational analysis. Experimental considerations include optimization of specimen preparation, objective acquisition of a sufficient number of spectra, linking of the derived spectra with tissue architecture or cell type, and computational analysis. The preparation of multiple specimens (up to 50) takes 8 h; the interrogation of a tissue section can take up to 6 h (similar to 100 spectra); and cytology analysis (n = 50, 10 spectra per specimen) takes 14 h. IR spectroscopy generates complex data sets and analyses are best when initially based on a multivariate approach (principal component analysis with or without linear discriminant analysis). This results in the identification of class clustering as well as class-specific chemical entities.