Multispectral imaging can be employed for investigation of paintings and other works of art. The painting is irradiated by ultraviolet, visible and infrared rays and the reflected radiation is recorded in a camera sensitive in this region of the spectrum. The image can also be registered using the transmitted instead of reflected radiation. In special cases the painting can be irradiated by UV, VIS or IR rays and the fluorescence of pigments or varnishes can be registered.
Multispectral analysis has assisted in the interpretation of ancient papyri, such as those found at Herculaneum, by imaging the fragments in the infrared range (1000 nm). Often, the text on the documents appears to the naked eye as black ink on black paper. At 1000 nm, the difference in how paper and ink reflect infrared light makes the text clearly readable. It has also been used to image the Archimedes palimpsest by imaging the parchment leaves in bandwidths from 365–870 nm, and then using advanced digital image processing techniques to reveal the undertext with Archimedes' work. Multispectral imaging has been used in a Mellon Foundation project at Yale University to compare inks in medieval English manuscripts.
Multispectral imaging has also been used to examine discolorations and stains on old books and manuscripts.  Comparing the "spectral fingerprint" of a stain to the characteristics of known chemical substances can make it possible to identify the stain. This technique has been used to examine medical and alchemical texts, seeking hints about the activities of early chemists and the possible chemical substances they may have used in their experiments. Like a cook spilling flour or vinegar on a cookbook, an early chemist might have left tangible evidence on the pages of the ingredients used to make medicines.