Foraminiferal, algal and problematica assemblages from the Mississippian (late Viséan and early Serpukhovian) Lower Limestone Formation have been studied in order to validate lithostratigraphical correlations of limestones within the central and western parts of the Midland Valley of Scotland. Analysis of more than 100 outcrops allows recognition of four calcareous microfossil assemblages, which span the late Brigantian and early Pendleian, and enables a detailed correlation to be made within the Central Coalfield (north Lanarkshire) and with the thinner sequences to the west (north Ayrshire), to the south (Douglas area, south Lanarkshire), and to the east (Bathgate area, West Lothian). The age of the Lower Limestone Formation is modified because the upper part of this formation is now assigned to the Pendleian (due to the first occurrences of new foraminiferans and the co-occurrence with the Namurian goniatites), and some individual limestone horizons within the formation are repositioned, or their precise correlation with other limestones is established. A refined stratigraphical framework is proposed for the above noted areas, and a correlation between them and the Pennine region in northern England is proposed, passing through the Archerbeck Borehole sequence in the Scottish Borders.