Self-supervised Pretraining of Cell Segmentation Models

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.

Accurate delineation of individual cells in microscopy videos is essential for studying cellular dynamics, yet separating touching or overlapping instances remains a persistent challenge. Although foundation-model for segmentation such as SAM have broadened the accessibility of image segmentation, they still struggle to separate nearby cell instances in dense microscopy scenes without extensive prompting. We propose a prompt-free, boundary-aware instance segmentation framework that predicts signed distance functions (SDFs) instead of binary masks, enabling smooth and geometry-consistent modeling of cell contours. A learned sigmoid mapping converts SDFs into probability maps, yielding sharp boundary localization and robust separation of adjacent instances. Training is guided by a unified Modified Hausdorff Distance (MHD) loss that integrates region- and boundary-based terms. Evaluations on both public and private high-throughput microscopy datasets demonstrate improved boundary accuracy and instance-level performance compared to recent SAM-based and foundation-model approaches. Source code is available at: https://github.com/ThomasMendelson/BAISeg.git

Background: Task-specific microscopy datasets are often small, making it difficult to train deep learning models that learn robust features. While self-supervised learning (SSL) has shown promise through pretraining on large, domain-specific datasets, generalizability across datasets with differing staining protocols and channel configurations remains underexplored. We investigated the generalizability of SSL models pretrained on ImageNet-1k and HPA FOV, evaluating their embeddings on OpenCell with and without fine-tuning, two channel-mismatch strategies, and varying fine-tuning data fractions. We additionally analyzed single-cell embeddings on a labeled OpenCell subset. Result: DINO-based ViT backbones pretrained on HPA FOV or ImageNet-1k transfer well to OpenCell even without fine-tuning. Fine-tuning further improved performance to 0.704 $\pm$ 0.027 (17 classes, multi-class labels). At the single-cell level, the HPA single-cell-pretrained model achieved the highest k-nearest neighbor performance across all neighborhood sizes (macro $F_1$ $\geq$ 0.515). Conclusion: SSL methods like DINO, pretrained on large domain-relevant datasets, enable effective use of deep learning features for fine-tuning on small, task-specific microscopy datasets.