The Middle Marker – horizon H1 of the Hooggenoeg Formation – is the oldest sedimentary horizon in the Barberton greenstone belt and one of the oldest sedimentary horizons on Earth. Herein, we describe a range of carbonaceous microstructures in this unit which bear resemblance to phototrophic microbial biofilms, biosedimentary structures, and interpreted microfossils in contemporaneous greenstone belts from the Early Archaean. Post-depositional iron-rich fluid cycling through these sediments has resulted in the precipitation of pseudo-laminated structures, which also bear resemblance, at the micron-scale, to certain microbial mat-like structures, although are certainly abiogenic. Poor preservation of multiple putative microbial horizons due to coarse volcaniclastic sedimentation and synsedimentary fragmentation by hydrothermal fluid also makes a conclusive assessment of biogenicity challenging. Nonetheless, several laminated morphologies within volcaniclastic sandstones and siltstones and coarse-grained volcaniclastic sandstones are recognisable as syngenetic photosynthetic microbial biofilms and microbially induced sedimentary structures; therefore, the Middle Marker preserves the oldest evidence for life in the Barberton greenstone belt. Among these biosignatures are fine, crinkly, micro-tufted, laminated microbial mats, pseudo-tufted laminations and wisp-like carbonaceous fragments interpreted as either partially formed biofilms or their erosional products. In the same sediments, lenticular objects, which have previously been interpreted as bona fide microfossils, are rare but recurrent finds whose biogenicity we question. The Middle Marker preserves an ancient record of epibenthic microbial communities flourishing, struggling and perishing in parallel with a waning volcanic cycle, an environment upon which they depended and through which they endured. Direct comparisons can be made between environment-level characters of the Middle Marker and other Early Archaean cherts, suggesting that shallow-water, platformal, volcanogenic-hydrothermal biocoenoses were major microbial habitats throughout the Archaean.