Abstract
BackgroundMicroscopy image segmentation lays the foundation for shape analysis, motion tracking, and classification of biological objects. Despite its importance, automated segmentation remains challenging for several widely used non-fluorescence, interference-based microscopy imaging modalities. For example in differential interference contrast microscopy which plays an important role in modern bacterial cell biology. Therefore, new revolutions in the field require the development of tools, technologies and work-flows to extract and exploit information from interference-based imaging data so as to achieve new fundamental biological insights and understanding.ResultsWe have developed and evaluated a high-throughput image analysis and processing approach to detect and characterize bacterial cells and chemotaxis proteins. Its performance was evaluated using differential interference contrast and fluorescence microscopy images of Rhodobacter sphaeroides.ConclusionsResults demonstrate that the proposed approach provides a fast and robust method for detection and analysis of spatial relationship between bacterial cells and their chemotaxis proteins.
Highlights
Microscopy image segmentation lays the foundation for shape analysis, motion tracking, and classification of biological objects
Generating contrast in these images to determine the cell boundary is achieved using a number of optical methods including Phase Contrast and Differential Interference Contrast (DIC) microscopy both of which depend on light changing its properties as it passes through the sample
It is increasingly important to interpret microscopy images in a quantitative manner being able to reconstruct the cell boundary from these images is of great importance, allowing the location of fluorescent markers to be determined within the bacterial cell
Summary
Microscopy image segmentation lays the foundation for shape analysis, motion tracking, and classification of biological objects. Modern bacterial cell biology has been revolutionised with the use of fluorescent markers coupled with microscopy allowing the visualisation of sub-cellular localisation in the bacterial cell. Generating contrast in these images to determine the cell boundary is achieved using a number of optical methods including Phase Contrast and Differential Interference Contrast (DIC) microscopy both of which depend on light changing its properties as it passes through the sample. It is increasingly important to interpret microscopy images in a quantitative manner being able to reconstruct the cell boundary from these images is of great importance, allowing the location of fluorescent markers to be determined within the bacterial cell. In the perpendicular direction to the DIC shear there is no contrast against the background, and a lack of information about the complete cell boundary
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