In nature, bacteria are predominantly found as surface-associated cell assemblages known as biofilms.

In biofilms, bacteria do not become fundamentally different just in terms of their physiology and gene expression, they also acquire high resistance to antibiotics and a variety of antimicrobials.

Although the existence of biofilms has been documented for years, only recently have powerful tools for investigating them in situ been developed. One such tool is imaging using confocal laser scanning microscopy (CLSM) in combination with specific fluorescent stains.

CLSM explores the distribution and structure of biofilms in 2-D and 3-D, producing relatively large amounts of high resolution image data.

The amount of data acquired by CLSM calls for automatic image analysis methods which are both accurate and time-saving, in order that the visual information can be transformed into numbers.

bioImage_L is a novel image analysis software for the automatic characterization of the structure and distribution of biofilms. The novel feature of bioImage_L is the inclusion of an algorithm that automatically identifies color tonalities without having to make a prior monochrome conversion into separate RGB channels.