Quantitative tracking of tumor cells in phase-contrast microscopy exploiting halo artifact pattern
2012
Tumor cell morphology is closely related to its invasiveness characteristics and migratory behaviors. An invasive tumor
cell has a highly irregular shape, whereas a spherical cell is non-metastatic. Thus, quantitative analysis of cell features is
crucial to determine tumor malignancy or to test the efficacy of anticancer treatment. We use phase-contrast microscopy
to analyze single cell morphology and to monitor its change because it enables observation of long-term activity of living
cells without photobleaching and phototoxicity, which is common in other fluorescence-labeled microscopy. Despite this
advantage, there are image-level drawbacks to phase-contrast microscopy, such as local light effect and contrast
interference ring, among others.
Thus, we first applied a local filter to compensate for non-uniform illumination. Then, we used intensity distribution
information to detect the cell boundary. In phase-contrast microscopy images, the cell normally appears as a dark region
surrounded by a bright halo. As the halo artifact around the cell body is minimal and has an asymmetric diffusion pattern,
we calculated the cross-sectional plane that intersected the center of each cell and was orthogonal to the first principal
axis. Then, we extracted the dark cell region by level set. However, a dense population of cultured cells still rendered
single-cell analysis difficult. Finally, we measured roundness and size to classify tumor cells into malignant and benign
groups. We validated segmentation accuracy by comparing our findings with manually obtained results.
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