Multi-directional Morphological Assessment of Single Bacterial Colonies Through Non-invasive Optical Imaging

A bacterium in bacterial colony is a basal component of bacterial studies and is therefore of considerable importance. The morphological characteristics of a single colony have been widely used as the standard to identify diverse bacterial species. However, the conventional methods for obtaining morphological information, such as microscopic techniques, cannot provide tomographic views. This study utilizes spectral-domain optical coherence tomography (SD-OCT) to observe both external and internal structures for single colonies. OCT imaging provided a clear top (projection) and lateral (cross-sectional) images, which can identify distinct features in the fifteen different bacterial species examined. Through the acquired OCT images, the elevation type in each genus was confirmed, and the quantitative measurements for morphology including a diameter, height, and elevation angle were achieved. Moreover, the OCT raw data was used to build three-dimensional images to observe outer appearances in all direction. As a result, the averaged angles of 7.92 degrees, 14.40 degrees, 6.78 degrees, and 11.92 degrees of the genus Hymenobacter, Spirosoma, Bacillus, and Deinococcus, respectively, were calculated. In conclusion, we have demonstrated that OCT is a highly feasible, non-invasive method for the identification of bacterial species, providing detailed morphological and visual information. It thus has strong potential for applications in bacteriology, as well as biomedical and food sciences.