Dataset Information

Automated cell segmentation in FIJI® using the DRAQ5 nuclear dye.

ABSTRACT:

Background

Image segmentation and quantification are essential steps in quantitative cellular analysis. In this work, we present a fast, customizable, and unsupervised cell segmentation method that is based solely on Fiji (is just ImageJ)®, one of the most commonly used open-source software packages for microscopy analysis. In our method, the "leaky" fluorescence from the DNA stain DRAQ5 is used for automated nucleus detection and 2D cell segmentation.

Results

Based on an evaluation with HeLa cells compared to human counting, our algorithm reached accuracy levels above 92% and sensitivity levels of 94%. 86% of the evaluated cells were segmented correctly, and the average intersection over union score of detected segmentation frames to manually segmented cells was above 0.83. Using this approach, we quantified changes in the projected cell area, circularity, and aspect ratio of THP-1 cells differentiating from monocytes to macrophages, observing significant cell growth and a transition from circular to elongated form. In a second application, we quantified changes in the projected cell area of CHO cells upon lowering the incubation temperature, a common stimulus to increase protein production in biotechnology applications, and found a stark decrease in cell area.

Conclusions

Our method is straightforward and easily applicable using our staining protocol. We believe this method will help other non-image processing specialists use microscopy for quantitative image analysis.

SUBMITTER: Schwendy M

PROVIDER: S-EPMC6339324 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Publications

Automated cell segmentation in FIJI® using the DRAQ5 nuclear dye.

Schwendy Mischa M Unger Ronald E RE Bonn Mischa M Parekh Sapun H SH

BMC bioinformatics 20190118 1

<h4>Background</h4>Image segmentation and quantification are essential steps in quantitative cellular analysis. In this work, we present a fast, customizable, and unsupervised cell segmentation method that is based solely on Fiji (is just ImageJ)®, one of the most commonly used open-source software packages for microscopy analysis. In our method, the "leaky" fluorescence from the DNA stain DRAQ5 is used for automated nucleus detection and 2D cell segmentation.<h4>Results</h4>Based on an evaluati ...[more]

PMID: 30658582

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Project description:PurposeTo develop a fast and accurate convolutional neural network based method for segmentation of thalamic nuclei.MethodsA cascaded multi-planar scheme with a modified residual U-Net architecture was used to segment thalamic nuclei on conventional and white-matter-nulled (WMn) magnetization prepared rapid gradient echo (MPRAGE) data. A single network was optimized to work with images from healthy controls and patients with multiple sclerosis (MS) and essential tremor (ET), acquired at both 3 T and 7 T field strengths. WMn-MPRAGE images were manually delineated by a trained neuroradiologist using the Morel histological atlas as a guide to generate reference ground truth labels. Dice similarity coefficient and volume similarity index (VSI) were used to evaluate performance. Clinical utility was demonstrated by applying this method to study the effect of MS on thalamic nuclei atrophy.ResultsSegmentation of each thalamus into twelve nuclei was achieved in under a minute. For 7 T WMn-MPRAGE, the proposed method outperforms current state-of-the-art on patients with ET with statistically significant improvements in Dice for five nuclei (increase in the range of 0.05-0.18) and VSI for four nuclei (increase in the range of 0.05-0.19), while performing comparably for healthy and MS subjects. Dice and VSI achieved using 7 T WMn-MPRAGE data are comparable to those using 3 T WMn-MPRAGE data. For conventional MPRAGE, the proposed method shows a statistically significant Dice improvement in the range of 0.14-0.63 over FreeSurfer for all nuclei and disease types. Effect of noise on network performance shows robustness to images with SNR as low as half the baseline SNR. Atrophy of four thalamic nuclei and whole thalamus was observed for MS patients compared to healthy control subjects, after controlling for the effect of parallel imaging, intracranial volume, gender, and age (p < 0.004).ConclusionThe proposed segmentation method is fast, accurate, performs well across disease types and field strengths, and shows great potential for improving our understanding of thalamic nuclei involvement in neurological diseases.

Dataset Information

Automated cell segmentation in FIJI® using the DRAQ5 nuclear dye.

Background

Results

Conclusions

Publications

Automated cell segmentation in FIJI® using the DRAQ5 nuclear dye.

Similar Datasets

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