Constantin Pape

Matthias Eisenmann, Annika Reinke, Vivienn Weru et al. · utoronto

The number of international benchmarking competitions is steadily increasing in various fields of machine learning (ML) research and practice. So far, however, little is known about the common practice as well as bottlenecks faced by the community in tackling the research questions posed. To shed light on the status quo of algorithm development in the specific field of biomedical imaging analysis, we designed an international survey that was issued to all participants of challenges conducted in conjunction with the IEEE ISBI 2021 and MICCAI 2021 conferences (80 competitions in total). The survey covered participants' expertise and working environments, their chosen strategies, as well as algorithm characteristics. A median of 72% challenge participants took part in the survey. According to our results, knowledge exchange was the primary incentive (70%) for participation, while the reception of prize money played only a minor role (16%). While a median of 80 working hours was spent on method development, a large portion of participants stated that they did not have enough time for method development (32%). 25% perceived the infrastructure to be a bottleneck. Overall, 94% of all solutions were deep learning-based. Of these, 84% were based on standard architectures. 43% of the respondents reported that the data samples (e.g., images) were too large to be processed at once. This was most commonly addressed by patch-based training (69%), downsampling (37%), and solving 3D analysis tasks as a series of 2D tasks. K-fold cross-validation on the training set was performed by only 37% of the participants and only 50% of the participants performed ensembling based on multiple identical models (61%) or heterogeneous models (39%). 48% of the respondents applied postprocessing steps.

Anwai Archit, Constantin Pape

Segmentation is a crucial analysis task in biomedical imaging. Given the diverse experimental settings in this field, the lack of generalization limits the use of deep learning in practice. Domain adaptation is a promising remedy: it involves training a model for a given task on a source dataset with labels and adapts it to a target dataset without additional labels. We introduce a probabilistic domain adaptation method, building on self-training approaches and the Probabilistic UNet. We use the latter to sample multiple segmentation hypotheses to implement better pseudo-label filtering. We further study joint and separate source-target training strategies and evaluate our method on three challenging domain adaptation tasks for biomedical segmentation. Our code is publicly available at https://github.com/computational-cell-analytics/Probabilistic-Domain-Adaptation.

Anwai Archit, Constantin Pape

Cell segmentation is a fundamental task in microscopy image analysis. Several foundation models for cell segmentation have been introduced, virtually all of them are extensions of Segment Anything Model (SAM), improving it for microscopy data. Recently, SAM2 and SAM3 have been published, further improving and extending the capabilities of general-purpose segmentation foundation models. Here, we comprehensively evaluate foundation models for cell segmentation (CellPoseSAM, CellSAM, $μ$SAM) and for general-purpose segmentation (SAM, SAM2, SAM3) on a diverse set of (light) microscopy datasets, for tasks including cell, nucleus and organoid segmentation. Furthermore, we introduce a new instance segmentation strategy called automatic prompt generation (APG) that can be used to further improve SAM-based microscopy foundation models. APG consistently improves segmentation results for $μ$SAM, which is used as the base model, and is competitive with the state-of-the-art model CellPoseSAM. Moreover, our work provides important lessons for adaptation strategies of SAM-style models to microscopy and provides a strategy for creating even more powerful microscopy foundation models. Our code is publicly available at https://github.com/computational-cell-analytics/micro-sam.

Anwai Archit, Constantin Pape

CNNs, most notably the UNet, are the default architecture for biomedical segmentation. Transformer-based approaches, such as UNETR, have been proposed to replace them, benefiting from a global field of view, but suffering from larger runtimes and higher parameter counts. The recent Vision Mamba architecture offers a compelling alternative to transformers, also providing a global field of view, but at higher efficiency. Here, we introduce ViM-UNet, a novel segmentation architecture based on it and compare it to UNet and UNETR for two challenging microscopy instance segmentation tasks. We find that it performs similarly or better than UNet, depending on the task, and outperforms UNETR while being more efficient. Our code is open source and documented at https://github.com/constantinpape/torch-em/blob/main/vimunet.md.

Carolin Teuber, Anwai Archit, Tobias Boothe et al.

Deep learning underlies most modern approaches and tools in computer vision, including biomedical imaging. However, for interactive semantic segmentation (often called pixel classification in this context) and interactive object-level classification (object classification), feature-based shallow learning remains widely used. This is due to the diversity of data in this domain, the lack of large pretraining datasets, and the need for computational and label efficiency. In contrast, state-of-the-art tools for many other vision tasks in microscopy - most notably cellular instance segmentation - already rely on deep learning and have recently benefited substantially from vision foundation models (VFMs), particularly SAM. Here, we investigate whether VFMs can also improve pixel and object classification compared to current approaches. To this end, we evaluate several VFMs, including general-purpose models (SAM, SAM2, DINOv3) and domain-specific ones ($μ$SAM, PathoSAM), in combination with shallow learning and attentive probing on five diverse and challenging datasets. Our results demonstrate consistent improvements over hand-crafted features and provide a clear pathway toward practical improvements. Furthermore, our study establishes a benchmark for VFMs in microscopy and informs future developments in this area.

Anwai Archit, Luca Freckmann, Constantin Pape

Medical image segmentation is an important analysis task in clinical practice and research. Deep learning has massively advanced the field, but current approaches are mostly based on models trained for a specific task. Training such models or adapting them to a new condition is costly due to the need for (manually) labeled data. The emergence of vision foundation models, especially Segment Anything, offers a path to universal segmentation for medical images, overcoming these issues. Here, we study how to improve Segment Anything for medical images by comparing different finetuning strategies on a large and diverse dataset. We evaluate the finetuned models on a wide range of interactive and (automatic) semantic segmentation tasks. We find that the performance can be clearly improved for interactive segmentation. However, semantic segmentation does not benefit from pretraining on medical images. Our best model, MedicoSAM, is publicly available at https://github.com/computational-cell-analytics/medico-sam. We show that it is compatible with existing tools for data annotation and believe that it will be of great practical value.

Titus Griebel, Anwai Archit, Constantin Pape

Nucleus segmentation is an important analysis task in digital pathology. However, methods for automatic segmentation often struggle with new data from a different distribution, requiring users to manually annotate nuclei and retrain data-specific models. Vision foundation models (VFMs), such as the Segment Anything Model (SAM), offer a more robust alternative for automatic and interactive segmentation. Despite their success in natural images, a foundation model for nucleus segmentation in histopathology is still missing. Initial efforts to adapt SAM have shown some success, but did not yet introduce a comprehensive model for diverse segmentation tasks. To close this gap, we introduce PathoSAM, a VFM for nucleus segmentation, based on training SAM on a diverse dataset. Our extensive experiments show that it is the new state-of-the-art model for automatic and interactive nucleus instance segmentation in histopathology. We also demonstrate how it can be adapted for other segmentation tasks, including semantic nucleus segmentation. For this task, we show that it yields results better than popular methods, while not yet beating the state-of-the-art, CellViT. Our models are open-source and compatible with popular tools for data annotation. We also provide scripts for whole-slide image segmentation. Our code and models are publicly available at https://github.com/computational-cell-analytics/patho-sam.

Carolin Teuber, Anwai Archit, Constantin Pape

Segmentation is an important analysis task for biomedical images, enabling the study of individual organelles, cells or organs. Deep learning has massively improved segmentation methods, but challenges remain in generalization to new conditions, requiring costly data annotation. Vision foundation models, such as Segment Anything Model (SAM), address this issue through improved generalization. However, these models still require finetuning on annotated data, although with less annotations, to achieve optimal results for new conditions. As a downside, they require more computational resources. This makes parameter-efficient finetuning (PEFT) relevant. We contribute the first comprehensive study of PEFT for SAM applied to biomedical images. We find that the placement of PEFT layers is more important for efficiency than the type of layer for vision transformers and we provide a recipe for resource-efficient finetuning. Our code is publicly available at https://github.com/computational-cell-analytics/peft-sam.

Adrian Wolny, Qin Yu, Constantin Pape et al.

Most state-of-the-art instance segmentation methods have to be trained on densely annotated images. While difficult in general, this requirement is especially daunting for biomedical images, where domain expertise is often required for annotation and no large public data collections are available for pre-training. We propose to address the dense annotation bottleneck by introducing a proposal-free segmentation approach based on non-spatial embeddings, which exploits the structure of the learned embedding space to extract individual instances in a differentiable way. The segmentation loss can then be applied directly to instances and the overall pipeline can be trained in a fully- or weakly supervised manner. We consider the challenging case of positive-unlabeled supervision, where a novel self-supervised consistency loss is introduced for the unlabeled parts of the training data. We evaluate the proposed method on 2D and 3D segmentation problems in different microscopy modalities as well as on the Cityscapes and CVPPP instance segmentation benchmarks, achieving state-of-the-art results on the latter. The code is available at: https://github.com/kreshuklab/spoco

Artem Lukoyanov, Isabella Haberbosch, Constantin Pape et al.

Recent advances in high-throughput electron microscopy imaging enable detailed study of centrosome aberrations in cancer cells. While the image acquisition in such pipelines is automated, manual detection of centrioles is still necessary to select cells for re-imaging at higher magnification. In this contribution we propose an algorithm which performs this step automatically and with high accuracy. From the image labels produced by human experts and a 3D model of a centriole we construct an additional training set with patch-level labels. A two-level DenseNet is trained on the hybrid training data with synthetic patches and real images, achieving much better results on real patient data than training only at the image-level. The code can be found at https://github.com/kreshuklab/centriole_detection.

Larissa Heinrich, Jan Funke, Constantin Pape et al.

Neural circuit reconstruction at single synapse resolution is increasingly recognized as crucially important to decipher the function of biological nervous systems. Volume electron microscopy in serial transmission or scanning mode has been demonstrated to provide the necessary resolution to segment or trace all neurites and to annotate all synaptic connections. Automatic annotation of synaptic connections has been done successfully in near isotropic electron microscopy of vertebrate model organisms. Results on non-isotropic data in insect models, however, are not yet on par with human annotation. We designed a new 3D-U-Net architecture to optimally represent isotropic fields of view in non-isotropic data. We used regression on a signed distance transform of manually annotated synaptic clefts of the CREMI challenge dataset to train this model and observed significant improvement over the state of the art. We developed open source software for optimized parallel prediction on very large volumetric datasets and applied our model to predict synaptic clefts in a 50 tera-voxels dataset of the complete Drosophila brain. Our model generalizes well to areas far away from where training data was available.

Stefan Dvoretskii, Anwai Archit, Constantin Pape et al.

Modern bioimage analysis approaches are data hungry, making it necessary for researchers to scavenge data beyond those collected within their (bio)imaging facilities. In addition to scale, bioimaging datasets must be accompanied with suitable, high-quality annotations and metadata. Although established data repositories such as the Image Data Resource (IDR) and BioImage Archive offer rich metadata, their contents typically cannot be directly consumed by image analysis tools without substantial data wrangling. Such a tedious assembly and conversion of (meta)data can account for a dedicated amount of time investment for researchers, hindering the development of more powerful analysis tools. Here, we introduce BioimageAIpub, a workflow that streamlines bioimaging data conversion, enabling a seamless upload to HuggingFace, a widely used platform for sharing machine learning datasets and models.

Elena Buglakova, Anwai Archit, Edoardo D'Imprima et al.

Segmentation of very large images is a common problem in microscopy, medical imaging or remote sensing. The problem is usually addressed by sliding window inference, which can theoretically lead to seamlessly stitched predictions. However, in practice many of the popular pipelines still suffer from tiling artifacts. We investigate the root cause of these issues and show that they stem from the normalization layers within the neural networks. We propose indicators to detect normalization issues and further explore the trade-offs between artifact-free and high-quality predictions, using three diverse microscopy datasets as examples. Finally, we propose to use BatchRenorm as the most suitable normalization strategy, which effectively removes tiling artifacts and enhances transfer performance, thereby improving the reusability of trained networks for new datasets.

Paul Hilt, Maedeh Zarvandi, Edgar Kaziakhmedov et al.

Instance segmentation is an important computer vision problem which remains challenging despite impressive recent advances due to deep learning-based methods. Given sufficient training data, fully supervised methods can yield excellent performance, but annotation of ground-truth data remains a major bottleneck, especially for biomedical applications where it has to be performed by domain experts. The amount of labels required can be drastically reduced by using rules derived from prior knowledge to guide the segmentation. However, these rules are in general not differentiable and thus cannot be used with existing methods. Here, we relax this requirement by using stateless actor critic reinforcement learning, which enables non-differentiable rewards. We formulate the instance segmentation problem as graph partitioning and the actor critic predicts the edge weights driven by the rewards, which are based on the conformity of segmented instances to high-level priors on object shape, position or size. The experiments on toy and real datasets demonstrate that we can achieve excellent performance without any direct supervision based only on a rich set of priors.

Alberto Bailoni, Constantin Pape, Steffen Wolf et al.

This work introduces a new proposal-free instance segmentation method that builds on single-instance segmentation masks predicted across the entire image in a sliding window style. In contrast to related approaches, our method concurrently predicts all masks, one for each pixel, and thus resolves any conflict jointly across the entire image. Specifically, predictions from overlapping masks are combined into edge weights of a signed graph that is subsequently partitioned to obtain all final instances concurrently. The result is a parameter-free method that is strongly robust to noise and prioritizes predictions with the highest consensus across overlapping masks. All masks are decoded from a low dimensional latent representation, which results in great memory savings strictly required for applications to large volumetric images. We test our method on the challenging CREMI 2016 neuron segmentation benchmark where it achieves competitive scores.

Steffen Wolf, Yuyan Li, Constantin Pape et al.

Semantic instance segmentation is the task of simultaneously partitioning an image into distinct segments while associating each pixel with a class label. In commonly used pipelines, segmentation and label assignment are solved separately since joint optimization is computationally expensive. We propose a greedy algorithm for joint graph partitioning and labeling derived from the efficient Mutex Watershed partitioning algorithm. It optimizes an objective function closely related to the Symmetric Multiway Cut objective and empirically shows efficient scaling behavior. Due to the algorithm's efficiency it can operate directly on pixels without prior over-segmentation of the image into superpixels. We evaluate the performance on the Cityscapes dataset (2D urban scenes) and on a 3D microscopy volume. In urban scenes, the proposed algorithm combined with current deep neural networks outperforms the strong baseline of `Panoptic Feature Pyramid Networks' by Kirillov et al. (2019). In the 3D electron microscopy images, we show explicitly that our joint formulation outperforms a separate optimization of the partitioning and labeling problems.

Alberto Bailoni, Constantin Pape, Nathan Hütsch et al.

We propose a theoretical framework that generalizes simple and fast algorithms for hierarchical agglomerative clustering to weighted graphs with both attractive and repulsive interactions between the nodes. This framework defines GASP, a Generalized Algorithm for Signed graph Partitioning, and allows us to explore many combinations of different linkage criteria and cannot-link constraints. We prove the equivalence of existing clustering methods to some of those combinations and introduce new algorithms for combinations that have not been studied before. We study both theoretical and empirical properties of these combinations and prove that some of these define an ultrametric on the graph. We conduct a systematic comparison of various instantiations of GASP on a large variety of both synthetic and existing signed clustering problems, in terms of accuracy but also efficiency and robustness to noise. Lastly, we show that some of the algorithms included in our framework, when combined with the predictions from a CNN model, result in a simple bottom-up instance segmentation pipeline. Going all the way from pixels to final segments with a simple procedure, we achieve state-of-the-art accuracy on the CREMI 2016 EM segmentation benchmark without requiring domain-specific superpixels.

Constantin Pape, Alex Matskevych, Adrian Wolny et al.

The throughput of electron microscopes has increased significantly in recent years, enabling detailed analysis of cell morphology and ultrastructure. Analysis of neural circuits at single-synapse resolution remains the flagship target of this technique, but applications to cell and developmental biology are also starting to emerge at scale. The amount of data acquired in such studies makes manual instance segmentation, a fundamental step in many analysis pipelines, impossible. While automatic segmentation approaches have improved significantly thanks to the adoption of convolutional neural networks, their accuracy still lags behind human annotations and requires additional manual proof-reading. A major hindrance to further improvements is the limited field of view of the segmentation networks preventing them from exploiting the expected cell morphology or other prior biological knowledge which humans use to inform their segmentation decisions. In this contribution, we show how such domain-specific information can be leveraged by expressing it as long-range interactions in a graph partitioning problem known as the lifted multicut problem. Using this formulation, we demonstrate significant improvement in segmentation accuracy for three challenging EM segmentation problems from neuroscience and cell biology.

Steffen Wolf, Alberto Bailoni, Constantin Pape et al.

Image partitioning, or segmentation without semantics, is the task of decomposing an image into distinct segments, or equivalently to detect closed contours. Most prior work either requires seeds, one per segment; or a threshold; or formulates the task as multicut / correlation clustering, an NP-hard problem. Here, we propose an efficient algorithm for graph partitioning, the "Mutex Watershed''. Unlike seeded watershed, the algorithm can accommodate not only attractive but also repulsive cues, allowing it to find a previously unspecified number of segments without the need for explicit seeds or a tunable threshold. We also prove that this simple algorithm solves to global optimality an objective function that is intimately related to the multicut / correlation clustering integer linear programming formulation. The algorithm is deterministic, very simple to implement, and has empirically linearithmic complexity. When presented with short-range attractive and long-range repulsive cues from a deep neural network, the Mutex Watershed gives the best results currently known for the competitive ISBI 2012 EM segmentation benchmark.