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Cellular profiling with multiplexed immunofluorescence (MxIF) images can contribute to a more accurate patient stratification for immunotherapy. Accurate cell segmentation of the MxIF images is an essential step. We propose a deep learning pipeline to train a Mask R-CNN model (deep network) for cell segmentation using nuclear (DAPI) and membrane (Na + K + ATPase) stained images. We used two-stage domain adaptation by first using a weakly labeled dataset followed by fine-tuning with a manually annotated dataset. We validated our method against manual annotations on three different datasets. Our method yields comparable results to the multi-observer agreement on an ovarian cancer dataset and improves on state-of-the-art performance on a publicly available dataset of mouse pancreatic tissues. Our proposed method, using a weakly labeled dataset for pre-training, showed superior performance in all of our experiments. When using smaller training sample sizes for fine-tuning, the proposed method provided comparable performance to that obtained using much larger training sample sizes. Our results demonstrate that using two-stage domain adaptation with a weakly labeled dataset can effectively boost system performance, especially when using a small training sample size. We deployed the model as a plug-in to CellProfiler, a widely used software platform for cellular image analysis.

Cell segmentation for immunofluorescence multiplexed images using two-stage domain adaptation and weakly labeled data for pre-training