The Acidithiobacillus ferrooxidans response to stress associated with the drying process is known to be the production of extracellular polymeric substance (EPS) coverage. Here, samples of A. ferrooxidans suspensions grown in 1.8 pH and 3.0 pH and dried on mica and silicon are shown to form a structure of isolated bacteria. Individual bacteria coverage patterns were imaged by atomic force microscopy (AFM) on hydrophobic (silicon) and hydrophilic (mica) substrates. A comparison of images of covered and uncovered bacteria establish the volume of individual EPS coverage. The EPS production for bacteria on hydrophobic substrates shows a substantial decrease (a factor of 30) in the EPS volume per bacterium when compared with the one on hydrophilic substrates. Shape and volume determination of EPS structures on bacteria as a function of hydrophobicity or hydrophilicity of the substrate may help to determine the functions of EPS on bacterial aggregates.676312316