The present work exemplifies, over a mural painting from the 14th century, the advantages of an initial exhaustive research using latest generation hand-held spectrometers (Raman mainly) in order to perform the characterization of valuable objects of cultural heritage. These in-situ techniques (meaning on-site and non-destructive) are very useful to study the pigments and materials, to identify the nature and causes of some of the main sources of deterioration and to examine past repaints. In addition, the in-situ measurements are of great importance in the selection of micro-samples for the laboratory analyses. In this particular case, the combination of these results with the chemical imaging analyses in the laboratory (such as Raman and energy dispersive X-ray spectrometry imaging) allowed the characterization of the mural painting, including, the identification of all restoration works applied in the past. Copyright © 2014 John Wiley & Sons, Ltd.