Abstract:The quantitative analysis of microscope videos often requires instance segmentation and tracking of cellular and subcellular objects. The traditional method is composed of two stages: (1) performing instance object segmentation of each frame, and (2) associating objects frame-by-frame. Recently, pixel-embedding-based deep learning approaches provide single stage holistic solutions to tackle instance segmentation and tracking simultaneously. However, such deep learning methods require consistent annotations not only spatially (for segmentation), but also temporally (for tracking). In computer vision, annotated training data with consistent segmentation and tracking is resource intensive, the severity of which can be multiplied in microscopy imaging due to (1) dense objects (e.g., overlapping or touching), and (2) high dynamics (e.g., irregular motion and mitosis). To alleviate the lack of such annotations in dynamics scenes, adversarial simulations have provided successful solutions in computer vision, such as using simulated environments (e.g., computer games) to train real-world self-driving systems. In this paper, we propose an annotation-free synthetic instance segmentation and tracking (ASIST) method with adversarial simulation and single-stage pixel-embedding based learning. The contribution of this paper is three-fold: (1) the proposed method aggregates adversarial simulations and single-stage pixel-embedding based deep learning; (2) the method is assessed with both the cellular (i.e., HeLa cells) and subcellular (i.e., microvilli) objects; and (3) to the best of our knowledge, this is the first study to explore annotation-free instance segmentation and tracking study for microscope videos. This ASIST method achieved an important step forward, when compared with fully supervised approaches.
Abstract:Instance object segmentation and tracking provide comprehensive quantification of objects across microscope videos. The recent single-stage pixel-embedding based deep learning approach has shown its superior performance compared with "segment-then-associate" two-stage solutions. However, one major limitation of applying a supervised pixel-embedding based method to microscope videos is the resource-intensive manual labeling, which involves tracing hundreds of overlapped objects with their temporal associations across video frames. Inspired by the recent generative adversarial network (GAN) based annotation-free image segmentation, we propose a novel annotation-free synthetic instance segmentation and tracking (ASIST) algorithm for analyzing microscope videos of sub-cellular microvilli. The contributions of this paper are three-fold: (1) proposing a new annotation-free video analysis paradigm is proposed. (2) aggregating the embedding based instance segmentation and tracking with annotation-free synthetic learning as a holistic framework; and (3) to the best of our knowledge, this is first study to investigate microvilli instance segmentation and tracking using embedding based deep learning. From the experimental results, the proposed annotation-free method achieved superior performance compared with supervised learning.
Abstract:The unsupervised segmentation is an increasingly popular topic in biomedical image analysis. The basic idea is to approach the supervised segmentation task as an unsupervised synthesis problem, where the intensity images can be transferred to the annotation domain using cycle-consistent adversarial learning. The previous studies have shown that the macro-level (global distribution level) matching on the number of the objects (e.g., cells, tissues, protrusions etc.) between two domains resulted in better segmentation performance. However, no prior studies have exploited whether the unsupervised segmentation performance would be further improved when matching the exact number of objects at micro-level (mini-batch level). In this paper, we propose a deep learning based unsupervised segmentation method for segmenting highly overlapped and dynamic sub-cellular microvilli. With this challenging task, both micro-level and macro-level matching strategies were evaluated. To match the number of objects at the micro-level, the novel fluorescence-based micro-level matching approach was presented. From the experimental results, the micro-level matching did not improve the segmentation performance, compared with the simpler macro-level matching.