Reconstructing neuron morphologies from fluorescence microscope images plays a critical role in neuroscience studies. It relies on image segmentation to produce initial masks either for further processing or final results to represent neuronal morphologies. This has been a challenging step due to the variation and complexity of noisy intensity patterns in neuron images acquired from microscopes. Whereas progresses in deep learning have brought the goal of accurate segmentation much closer to reality, creating training data for producing powerful neural networks is often laborious. To overcome the difficulty of obtaining a vast number of annotated data, we propose a novel strategy of using two-stage generative models to simulate training data with voxel-level labels. Trained upon unlabeled data by optimizing a novel objective function of preserving predefined labels, the models are able to synthesize realistic 3D images with underlying voxel labels. We showed that these synthetic images could train segmentation networks to obtain even better performance than manually labeled data. To demonstrate an immediate impact of our work, we further showed that segmentation results produced by networks trained upon synthetic data could be used to improve existing neuron reconstruction methods.

Bioimage Informatics is a growing area that aims to extract biological knowledge from microscope images of biomedical samples automatically. Its mission is vastly challenging, however, due to the complexity of diverse imaging modalities and big scales of multi-dimensional images. One major challenge is automatic image segmentation, an essential step towards high-level modeling and analysis. While progresses in deep learning have brought the goal of automation much closer to reality, creating training data for producing powerful neural networks is often laborious. To provide a shortcut for this costly step, we propose a novel two-stage generative model for simulating voxel level training data based on a specially designed objective function of preserving foreground labels. Using segmenting neurons from LM (Light Microscopy) image stacks as a testing example, we showed that segmentation networks trained by our synthetic data were able to produce satisfactory results. Unlike other simulation methods available in the field, our method can be easily extended to many other applications because it does not involve sophisticated cell models and imaging mechanisms.

BACKGROUND: Normalization is a basic step in microarray data analysis. A proper normalization procedure ensures that the intensity ratios provide meaningful measures of relative expression values. METHODS: We propose a robust semiparametric method in a two-way semi-linear model (TW-SLM) for normalization of cDNA microarray data. This method does not make the usual assumptions underlying some of the existing methods. For example, it does not assume that: (i) the percentage of differentially expressed genes is small; or (ii) the numbers of up- and down-regulated genes are about the same, as required in the LOWESS normalization method. We conduct simulation studies to evaluate the proposed method and use a real data set from a specially designed microarray experiment to compare the performance of the proposed method with that of the LOWESS normalization approach. RESULTS: The simulation results show that the proposed method performs better than the LOWESS normalization method in terms of mean square errors for estimated gene effects. The results of analysis of the real data set also show that the proposed method yields more consistent results between the direct and the indirect comparisons and also can detect more differentially expressed genes than the LOWESS method. CONCLUSIONS: Our simulation studies and the real data example indicate that the proposed robust TW-SLM method works at least as well as the LOWESS method and works better when the underlying assumptions for the LOWESS method are not satisfied. Therefore, it is a powerful alternative to the existing normalization methods.