The recent development of Fourier Transform Infrared spectroscopic imaging devices provides a new imaging approach for cell and tissue pathology as distribution and structure of cellular components can be observed without sample staining. However, in order to translate this emerging technology to the clinic, optimization of the acquisition, classification and validation techniques is a prerequisite step. The information extracted from the spectral data largely depends on the quality of the raw spectra but also on the corrections and processing methods. In this chapter, some of the guidelines for the recording and preprocessing methods will be presented. Unsupervised statistical approaches for tissue type discrimination illustrate the potential of this technology on histopathology.