Abstract

BackgroundAnalysis of cellular processes with microscopic bright field defocused imaging has the advantage of low phototoxicity and minimal sample preparation. However bright field images lack the contrast and nuclei reporting available with florescent approaches and therefore present a challenge to methods that segment and track the live cells. Moreover, such methods must be robust to systemic and random noise, variability in experimental configuration, and the multiple unknowns in the biological system under study.ResultsA new method called maximal-information is introduced that applies a non-parametric information theoretic approach to segment bright field defocused images. The method utilizes a combinatorial optimization strategy to select specific defocused images from each image stack such that set complexity, a Kolmogorov complexity measure, is maximized. Differences among these selected images are then applied to initialize and guide a level set based segmentation algorithm. The performance of the method is compared with a recent approach that uses a fixed defocused image selection strategy over an image data set of embryonic kidney cells (HEK 293T) from multiple experiments. Results demonstrate that the adaptive maximal-information approach significantly improves precision and recall of segmentation over the diversity of data sets.ConclusionsIntegrating combinatorial optimization with non-parametric Kolmogorov complexity has been shown to be effective in extracting information from microscopic bright field defocused images. The approach is application independent and has the potential to be effective in processing a diversity of noisy and redundant high throughput biological data.

Highlights

  • Analysis of cellular processes with microscopic bright field defocused imaging has the advantage of low phototoxicity and minimal sample preparation

  • Cell segmentation is the identification of cell objects and their observable properties from biological images

  • Current cell segmentation methods perform most accurately when applied to high contrast and minimal noise images obtained from samples where the cells have fluorescentlylabeled cell nuclei and stained membranes, and are distinct with minimal adherent membranes

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Summary

Results

Set complexity analysis of image data To understand how Kolmogorov Complexity measures could reveal information in z-stacks, an initial study was performed by computing the NCD between each pair of 21 frames for three data sets each containing 192 z-stacks. In (b) maximal-information selects a alternative set of frames at different focus planes (compared to the fixed strategy) and produces significantly lower segmentation errors. In (b) maximal-information selects a alternative set of frames at different focus planes from the fixed strategy and produces significantly lower segmentation errors. Demonstrates the advantage of extracting more informative frames in the z-stack The average of both missing and unexpected cell segmentation for maximal-information are lower than sephaCe method. Availability and requirements Project name: maximal-information Project home page: https://sites.google.com/site/maxi malinformation, Operating system(s): Platform independent Programming language: Matlab Other requirements: requires sephaCe [3] downloaded from

Conclusions
Background
Motivation for the maximal information approach
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