Deep crustal felsic xenoliths from classic Scottish Midland Valley localities, carried to the surface by Permo-Carboniferous magmatism, are shown for the first time to include metaigneous varieties with dioritic and tonalitic protoliths. Four hypotheses regarding their origin have been evaluated: (1) Precambrian basement; (2) Permo-Carboniferous underplating; (3) ‘Newer Granite’ magmatism; (4) Ordovician arc magmatism. U–Pb zircon dating results rule out the Precambrian basement and Permo-Carboniferous underplating hypotheses, but establish that the meta-igneous xenoliths represent both ‘Newer Granite’ and Ordovician (to possibly Silurian) arc magmatism. The metadiorite xenoliths are shown to have protolith ages of c. 415 Ma with εHft zircon values ranging from +0·1 to +11·1. These are interpreted to represent unexposed ‘Newer Granite’ plutons, based on age, mineralogical, isotopic and geochemical data. This shows that Devonian ‘Newer Granite’ magmatism had a greater impact on the Midland Valley and Southern Uplands crust than previously realized. Clinopyroxene–plagioclase–quartz barometry on the metadiorites from the east and west of the Midland Valley yielded a similar pressure range of c. 5–10 kbar, and a metadiorite from the east yielded a minimum two-feldspar temperature estimate of c. 793–816°C. These results indicate that the metadiorites once resided in the middle–lower crust. In contrast, two metatonalite xenoliths have a Late Ordovician protolith age (c. 453 Ma), with zircon εHft values of +7·8 to +9·0. These are interpreted as samples of a buried Late Ordovician magmatic arc situated within the Midland Valley. Inherited zircons with similar Late Ordovician ages and εHft=453 values (+1·6 to +10·8) are present in the metadiorites, suggesting that the Devonian ‘Newer Granites’ intruded within or through this Late Ordovician Midland Valley arc. A younger protolith age of c. 430 Ma from one of the metatonalites suggests that arc activity continued until Silurian times. This validates the long-standing ‘arc collision’ hypothesis for the development of the Caledonian Orogen. Based on U–Pb zircon dating, the metatonalite and metadiorite xenoliths have both experienced metamorphism between c. 400 and c. 391 Ma, probably linked to the Acadian Orogeny. An older phase of metamorphism at c. 411 Ma was possibly triggered by the combined effects of heating owing to the emplacement of the ‘Newer Granite’ plutons and the overthrusting of the Southern Uplands terrane onto the southern margin of the Midland Valley terrane.