Scanning confocal laser microscopy (SCLM) technique is used in conjunction with analytical imaging techniques, digital analysis, and semi-automated image processing to assess the structure of biofilms and spatial variability within biofilms. The purpose of this study was to utilize digital images of biofilms from SCLM used in conjunction with fluorescent lectin probes to develop a tool to determine the volume occupied by bacteria in a certain region of the space and estimate of the number of bacteria. Applications may include the study of biofilm formation and growth, bacterial colonization, and determination of enzymatic activities.

Biofilms are formed by bacterial colonies encapsulated in an extra-cellular polymeric substances matrix. They form in short time if environmental conditions are favorable. The study of biofilms is facilitated by advances in microscopy, such as the SCLM technique used in conjunction with analytical imaging, digital analysis, and semi-automated image processing. Various approaches were developed based on SCLM to assess the structure of biofilms and spatial variability within biofilms. The objective is to utilize digital images of biofilms from SCLM used in conjunction with fluorescent lectin probes to develop a tool that will eventually determine the volume occupied by bacteria in a certain region of the space.