Segmentation and learning in the quantitative analysis of microscopy images

Semi-automated segmentation or more commonly known as interactive image segmentation is an algorithm that extracts a region of interest (ROI) from an image based on the input information from the user. The said algorithm will be repetitively fed with such input information until required region of interest is successfully segmented. To accelerate this segmentation procedure as well as enhancing the result, pre-processing steps can be applied. The application of superpixel is an example of such pre-processing step. Superpixel can be defined as a collection of pixels that share common features such as texture and colours. Though employed as pre-processing step in many interactive segmentation algorithms, to date, no study has been conducted to assess the effects of such incorporations on the segmentation algorithms. Thus, this study aims to address this issue. In this study, five different types of superpixels ranging from watershed, density, graph, clustering and energy optimization categories are evaluated. The superpixels generated by these five algorithms will be used on two interactive image segmentation algorithms: i) Maximal Similarity based Region Merging (MSRM) and ii) Graph-Based Manifold Ranking (GBMR) with single and multiple strokes on various images from the Berkeley image dataset. The result of testing had shown that MSRM achieved better result compared to GBMR in both single and multiple input strokes using SEEDS superpixel algorithm. This study summary concluded that at different superpixel algorithms produced different results and that it is not possible to single out one particular superpixel algorithm that can work well for all the interactive segmentation algorithms. As such, the key to achieving a decent segmentation result lies in choosing the right superpixel algorithms for a given interactive segmentation algorithm.

This study investigates the impact of image complexity on the accuracy of interactive image segmentation algorithms. Image complexity plays a crucial role in segmentation performance, yet previous studies have primarily relied on subjective methods, leaving a gap in understanding how objective measures impact accuracy. The purpose of this research is to explore the relationship between image complexity and segmentation performance and to propose an adaptive approach for improving accuracy based on complexity measures. The study utilizes objective measures, namely entropy and fractal dimension, to quantify image complexity. An interactive image segmentation algorithm is employed, with a bounding box as the background and strokes as the foreground annotations. The number of strokes is dynamically adjusted based on complexity measures, ensuring a tailored segmentation approach. Comparative evaluations are conducted to assess the effectiveness of dynamic and fixed stroke allocation strategies. The principal results reveal a significant influence of image complexity on segmentation accuracy. The dynamic stroke allocation strategy outperforms fixed allocation, highlighting the importance of adapting to complexity. Moreover, the optimal combination of strokes and superpixel sizes is explored, providing valuable insights for practitioners. The findings emphasize the need to consider image complexity in segmentation algorithms to achieve accurate results. In conclusion, this study contributes to the understanding of the relationship between image complexity and interactive image segmentation. By introducing a dynamic stroke allocation approach and evaluating different configurations, the research provides insights into optimizing accuracy based on image complexity. The adaptive strategy improves segmentation performance and guides the development of robust algorithms. Future research can further refine the adaptive approach, explore additional complexity measures, and incorporate advanced machine learning techniques to enhance interactive image segmentation. Overall, this study advances the field by highlighting the importance of image complexity, providing guidance for practitioners, and paving the way for more efficient segmentation algorithms.

Convolutional neural networks (CNNs) are the backbone of most modern interactive segmentation algorithms. However, the limited receptive field of CNNs restricts their ability to capture long-range semantic relationships. Recently, transformers have gained significant attention for their capacity to capture long-range dependencies. Nevertheless, CNNs still outperform Transformer in extracting local information. An effective interactive segmentation algorithm should accurately capture fine-grained local details alongside global semantic relationships. Therefore, we propose GLClick, a global–local click-based interactive image segmentation model that integrates local and global information through a novel fusion mechanism. We design an efficient global–local feature fusion module (GLFM) that integrates fine-grained features from various layers of ResNet50 with those from the Transformer feature pyramid. This approach maintains ResNet50’s ability to extract local features while effectively leveraging the Transformer to capture global context. Additionally, we enhance the multi-layer perceptron (MLP) to improve performance. Extensive experiments on diverse benchmark datasets demonstrate significant improvements in interactive image segmentation, confirming the effectiveness of our approach. Moreover, we conduct experiments on medical image datasets, further illustrating the model’s versatility and effectiveness across different domains.

Interactive segmentation methods utilize user-provided positive and negative clicks to guide the model in accurately segmenting target objects. Compared to fully automatic medical image segmentation, these methods can achieve higher segmentation accuracy with limited image data, demonstrating significant potential in clinical applications. Typically, for each new click provided by the user, conventional interactive segmentation methods reprocess the entire network by re-inputting the click into the segmentation model, which greatly increases the user’s interaction burden and deviates from the intended goal of interactive segmentation tasks. To address this issue, we propose an efficient segmentation network, ESM-Net, for interactive medical image segmentation. It obtains high-quality segmentation masks based on the user’s initial clicks, reducing the complexity of subsequent refinement steps. Recent studies have demonstrated the strong performance of the Mamba model in various vision tasks; however, its application in interactive segmentation remains unexplored. In our study, we incorporate the Mamba module into our framework for the first time and enhance its spatial representation capabilities by developing a Spatial Augmented Convolution (SAC) module. These components are combined as the fundamental building blocks of our network. Furthermore, we designed a novel and efficient segmentation head to fuse multi-scale features extracted from the encoder, optimizing the generation of the predicted segmentation masks. Through comprehensive experiments, our method achieved state-of-the-art performance on three medical image datasets. Specifically, we achieved 1.43 NoC@90 on the Kvasir-SEG dataset, 1.57 NoC@90 on the CVC-ClinicDB polyp segmentation dataset, and 1.03 NoC@90 on the ADAM retinal disk segmentation dataset. The assessments on these three medical image datasets highlight the effectiveness of our approach in interactive medical image segmentation.

Extensive research has been conducted in an effort to evaluate methods and techniques for image segmentation. However, while most literature has focused on evaluating automatic and semi-automatic algorithms, works evaluating interactive segmentation algorithms are less numerous. Note that interactive segmentation can improve results by adding prior knowledge from users into the process. Although this user guidance improves segmentation results, it also makes difficult to conduct objective evaluations. For this reason, some works only present non-canonical evaluations. In this paper, we present an objective and empirical evaluation of seed-based interactive segmentation algorithms. We first compare popular metrics that are employed in image-segmentation evaluations in order to define which one reflects most accurately the performance of segmentation algorithms. Then, in the aim of presenting reproducible results, we introduce a novel seed-based user input dataset that extends the well-known GrabCut dataset. In addition, we evaluate and contrast four state-of-the-art interactive segmentation algorithms. The analysis of the results demonstrates that Jaccard coefficient and Precision-Recall curves provide a good insight into the performance of the evaluated algorithms. Finally, the GrabCut algorithm presents the most robust and useful segmentation among all the evaluated algorithms.

Interactive Image Segmentation has attracted much attention in the vision and graphics community recently. A typical application for interactive image segmentation is foreground/background segmentation based on user specified brush labellings. The problem can be formulated within the binary Markov Random Field (MRF) framework which can be solved efficiently via graph cut [1]. However, no attempt has yet been made to handle segmentation of multiple regions using graph cuts. In this paper, we propose a multiclass interactive image segmentation algorithm based on the Potts MRF model. Following [2], this can be converted to a multiway cut problem first proposed in [2] and solved by expansion-move algorithms for approximate inference [2]. A faster algorithm is proposed in this paper for efficient solution of the multiway cut problem based on partial optimal labeling. To achieve this, we combine the one-vs-all classifier fusion framework with the expansion-move algorithm for label inference over large images. We justify our approach with both theoretical analysis and experimental validation.

Interactive image segmentation based on samples reconstruction and FLDA

MotivationOver the last decades, image processing and analysis have become one of the key technologies in systems biology and medicine. The quantification of anatomical structures and dynamic processes in living systems is essential for understanding the complex underlying mechanisms and allows, i.e. the construction of spatio-temporal models that illuminate the interplay between architecture and function. Recently, deep learning significantly improved the performance of traditional image analysis in cases where imaging techniques provide large amounts of data. However, if only a few images are available or qualified annotations are expensive to produce, the applicability of deep learning is still limited.ResultsWe present a novel approach that combines machine learning-based interactive image segmentation using supervoxels with a clustering method for the automated identification of similarly colored images in large image sets which enables a guided reuse of interactively trained classifiers. Our approach solves the problem of deteriorated segmentation and quantification accuracy when reusing trained classifiers which is due to significant color variability prevalent and often unavoidable in biological and medical images. This increase in efficiency improves the suitability of interactive segmentation for larger image sets, enabling efficient quantification or the rapid generation of training data for deep learning with minimal effort. The presented methods are applicable for almost any image type and represent a useful tool for image analysis tasks in general.Availability and implementationThe presented methods are implemented in our image processing software TiQuant which is freely available at tiquant.hoehme.com.Supplementary informationSupplementary data are available at Bioinformatics online.

Interactive image segmentation is a challenging task and receives increasing attention recently; however, two major drawbacks exist in interactive segmentation approaches. First, the segmentation performance of region of interest (ROI)-based methods is sensitive to the initial ROI: different ROIs may produce results with great difference. Second, most seed-based methods need intense interactions, and are not applicable in many cases. In this paper, we generalize the neutro-connectedness (NC) to be independent of top-down priors of objects and to model image topology with indeterminacy measurement on image regions, propose a novel method for determining object and background regions, which is applied to exclude isolated background regions and enforce label consistency, and put forward a hybrid interactive segmentation method, NC Cut (NC-Cut), which can overcome the above two problems by utilizing both pixelwise appearance information and region-based NC properties. We evaluate the proposed NC-Cut by employing two image data sets (265 images), and demonstrate that the proposed approach outperforms the state-of-the-art interactive image segmentation methods (Grabcut, MILCut, One-Cut, MGCmaxsum, and pPBC).

Interactive image segmentation enables users to interact minimally with a machine, facilitating the gradual refinement of the segmentation mask for a target of interest. Previous studies have demonstrated impressive performance in extracting a single target mask through interactive segmentation. However, existing methods often overlook the information cues from previously interacted objects, despite realworld scenarios frequently involving multiple objects in the same category, leading to increased redundant user effort. To this end, this chapter introduces a new interactive image segmentation framework for multiple objects within the same category in a single image. Specifically, this work leverages Transformers to extract interactionfocused visual features from the image and the interactions, thereby obtaining a satisfactory mask of the target as an exemplar. For multiple objects, an exemplar -informed module is proposed to enhance the learning of similarities among the objects of the target category. To combine features from different modules, cross-attention blocks are incorporated, followed by a feature fusion module. Extensive experiments conducted on different datasets demonstrate that the proposed framework achieves superior performance compared to previous methods. Particularly, it reduces users’ labor by around 15%, requiring two fewer clicks to achieve target IoUs of 85% and 90% for multiple objects. These results further highlight the proposed method’s potential as a flexible and practical annotation tool.

Interactive segmentation has recently been explored to effectively and efficiently harvest high-quality segmentation masks by iteratively incorporating user hints. While iterative in nature, most existing interactive segmentation methods tend to ignore the dynamics of successive interactions and take each interaction independently. We here propose to model iterative interactive image segmentation with a Markov decision process (MDP) and solve it with reinforcement learning (RL) where each voxel is treated as an agent. Considering the large exploration space for voxel-wise prediction and the dependence among neighboring voxels for the segmentation tasks, multi-agent reinforcement learning is adopted, where the voxel-level policy is shared among agents. Considering that boundary voxels are more important for segmentation, we further introduce a boundary-aware reward, which consists of a global reward in the form of relative cross-entropy gain, to update the policy in a constrained direction, and a boundary reward in the form of relative weight, to emphasize the correctness of boundary predictions. To combine the advantages of different types of interactions, i. e., simple and efficient for point-clicking, and stable and robust for scribbles, we propose a supervoxel-clicking based interaction design. Experimental results on four benchmark datasets have shown that the proposed method significantly outperforms the state-of-the-arts, with the advantage of fewer interactions, higher accuracy, and enhanced robustness.

The image segmentation of histopathological tissue images has always been a challenge due to the overlapping of tissue color distributions, the complexity of extracellular texture and the large image size. In this paper, we introduce a new region-merging algorithm, namely, the Regional Pattern Merging (RPM) for interactive color image segmentation and annotation, by efficiently retrieving and applying the user’s prior knowledge of stroke-based interaction. Low-level color/texture features of each region are used to compose a regional pattern adapted to differentiating a foreground object from the background scene. This iterative region-merging is based on a modified Region Adjacency Graph (RAG) model built from initial segmented results of the mean shift to speed up the merging process. The foreground region of interest (ROI) is segmented by the reduction of the background region and discrimination of uncertain regions. We then compare our method against state-of-the-art interactive image segmentation algorithms in both natural images and histological images. Taking into account the homogeneity of both color and texture, the resulting semi-supervised classification and interactive segmentation capture histological structures more completely than other intensity or color-based methods. Experimental results show that the merging of the RAG model runs in a linear time according to the number of graph edges, which is essentially faster than both traditional graph-based and region-based methods.

Nuclear forensics relies on different signatures to identify the source of nuclear material. Such signatures include crystalline structure, chemical composition, and particle morphology. One way to quantify morphology in electron microscope imagery is through image segmentation, where pixels are assigned to several partitions (or groups) that correspond to particles, grains, and other objects of interest within the image. Once pixels are assigned to segments, it is relatively straightforward to quantify other quantities of interest, such as grain size, circularity, etc. However, the range and diversity of microscope images make it difficult to obtain an accurate segmentation automatically. The accuracy of segmentation can be improved through supervised learning, but this requires many images to be manually segmented. Another way to improve the accuracy is to use interactive segmentation. Interactive segmentation requires a human to provide image-specific user input to improve performance. However, the amount of user input (effort) is generally far less than is required for supervised learning. In this paper, we investigate several parallelization strategies to automatically explore the user input parameter space of interactive segmentation algorithms across a large number of images. Specifically, we investigate four different parallelization mechanisms in a high-performance computing (HPC) environment and use the Amdahl fraction to evaluate efficiency on multiple processor cores across multiple nodes. Ultimately, the parallelization strategy that was most efficient utilized the message passing interface integrated with the segmentation and quantification code. This strategy had an Amdahl fraction of 0.985 and a performance of about 0.251 s/image. These results indicate that the parameter space of interactive segmentation algorithms can be efficiently explored using HPC. This opens the door to future work where user input is reduced and where interactive image segmentation algorithms are automatically applied to large image sets.

A new algorithm for interactive image segmentation is proposed. Besides the traditional appearance and gradient information, a new Generic Shape Prior (GSP) knowledge which implies the location and the shape information of the object is combined into the framework. The GSP can be further categorized into the Regional and the Contour GSP to fit the interactive application, where a hierarchical graph-cut based optimization procedure is established, for its global optimization using the regional GSP to obtain good global segmentation results, and the local one using the Contour GSP to refine boundaries of global results. Moreover, the global optimization is based on superpixels which significantly reduce the computational complexity but preserve necessary image structures; the local one only considers a subset pixels around a contour segment, they both speed up the system. Results show our method performs better on both speed and accuracy.

This paper proposes an algorithm for interactive image segmentation. The task is formulated as a problem of graph-based transductive classification. Specifically, given an image window, the color of each pixel in it will be reconstructed linearly with those of the remaining pixels in this window. The optimal reconstruction weights will be kept unchanged to linearly reconstruct their class labels. The label reconstruction errors are estimated in each window. These errors are further collected together to develop a learning model. Then, the class information about the user specified foreground and background pixels are integrated into a regularization framework. Under this framework, a globally optimal labeling is finally obtained. The computational complexity is analyzed, and an approach for speeding up the algorithm is presented. Comparative experimental results illustrate the validity of our algorithm.