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Abstract: This paper presents a new investigation of the Portrait of Paul V Borghese, whose recent inclusion in the exhibition Caravaggio a Roma. Una vita dal vero has given new vigour to the debate over its attribution to Caravaggio. A series of considerations through stylistic comparisons dispute the alternative authorships that have been put forward for the painting, it then presents the results of studies carried out in the State Archives of Rome and the Secret Vatican Archives. These documents, cross-referenced with biographical sources and inventories belonging to the Borghese family, form the basis of a proposed reconstruction of the historical circumstances which led to the creation of the portrait, of the place in which it was painted, of its conservation and of the attribution to Caravaggio. These propositions are supported by findings that have emerged from laboratory analyses carried out on the painting. Radiography, reflectography, macrophotography and fluorescent imagery have revealed the presence of scratches and various pentimenti on the face of the Pope, as well as showing the presence of two separate layers of paint, suggesting that another painter may have worked at a later date and in a different style. An examination of the scratches on the face and of the overall chiaroscuro has been combined with an analysis of the reflection depicted on the spherical extremity of the seat’s backrest. This detail, which until now has not been mentioned, is compatible with the light which would have been cast from the Loggia d’Onore of the Quirinal Palace into the the ‘Camera del Bronzino’ at midday at the beginning of the 17th century, making it very probable that the Pope sat for his portrait in this very room.