Microalgal biofilms are associated with considerable variability in the properties of natural sediments, yet little effort has been made to isolate micro-scale spatial and temporal changes in sediment properties caused by the growth of a biofilm. Understanding the changes associated with biofilm growth and quantifying the time scales over which these changes occur is important for developing suitable experimental designs and for understanding how biofilms mediate sediment properties and processes. The development of a microphytobenthic biofilm and associated changes in the sediment was investigated over 45 days in the laboratory. The biogeochemical properties of the sediment: bulk density, water content, chlorophyll a concentration and colloidal carbohydrate concentration were measured on a sub-millimetre scale in the top 2 mm. The erosion threshold was measured with a Cohesive Strength Meter (CSM). Biofilm development was rapid, with changes in the properties occurring after 1 day and a visible film forming after just 3 days. The largest changes in sediment properties tended to occur in the surface 200 mm through time, with some variables also showing a differing response with depth. There were significant changes in water content, chlorophyll a concentration, colloidal carbohydrate concentration and erosion threshold in the surface 2 mm, with a general trend to increase with time. Bulk density was highly variable and did not show a consistent pattern of change with time. Erosion threshold was positively correlated with water content, chlorophyll a and colloidal carbohydrate in the surface 200 mm and these were also positively correlated with each other. Low Temperature Scanning Electron Microscopy (LTSEM) images revealed changes in the surface sediment structure and the formation of a thick multi-layer biofilm. The rapidity of biofilm growth and development and the associated changes to the sediment should be considered when designing experiments that investigate biofilms and properties of sediments and/or that involve biocide treatments or disturbance to the sediment.