Recent deep learning methods have provided successful initial segmentation results for generalized cell segmentation in microscopy. However, for dense arrangements of small cells with limited ground truth for training, the deep learning methods produce both over-segmentation and under-segmentation errors. Post-processing attempts to balance the trade-off between the global goal of cell counting for instance segmentation, and local fidelity to the morphology of identified cells. The need for post-processing is especially evident for segmenting 3D bacterial cells in densely-packed communities called biofilms. A graph-based recursive clustering approach, m-LCuts, is proposed to automatically detect collinearly structured clusters and applied to post-process unsolved cells in 3D bacterial biofilm segmentation. Construction of outlier-removed graphs to extract the collinearity feature in the data adds additional novelty to m-LCuts. The superiority of m-LCuts is observed by the evaluation in cell counting with over 90% of cells correctly identified, while a lower bound of 0.8 in terms of average single-cell segmentation accuracy is maintained. This proposed method does not need manual specification of the number of cells to be segmented. Furthermore, the broad adaptation for working on various applications, with the presence of data collinearity, also makes m-LCuts stand out from the other approaches.

Bacterial biofilm segmentation poses significant challenges due to lack of apparent structure, poor imaging resolution, limited contrast between conterminous cells and high density of cells that overlap. Although there exist bacterial segmentation algorithms in the existing art, they fail to delineate cells in dense biofilms, especially in 3D imaging scenarios in which the cells are growing and subdividing in a complex manner. A graph-based data clustering method, £Cuts, is presented with the application on bacterial cell segmentation. By constructing a weighted graph with node features in locations and principal orientations, the proposed method can automatically classify and detect differently oriented aggregations of linear structures (represent by bacteria in the application). The method assists in the assessment of several facets, such as bacterium tracking, cluster growth, and mapping of migration patterns of bacterial biofilms. Quantitative and qualitative measures for 2D data demonstrate the superiority of proposed method over the state of the art. Preliminary 3D results exhibit reliable classification of the cells with 97% accuracy.