Hélène Bouvrais

Sidi Mohamed Sid El Moctar, Achraf Ait Laydi, Yousef El Mourabit et al.

Microtubules are cytoskeletal filaments that play essential roles in many cellular processes and are key therapeutic targets in several diseases. Accurate segmentation of microtubule networks is critical for studying their organization and dynamics but remains challenging due to filament curvature, dense crossings, and image noise. We present MTFlow, a novel time-conditioned flow-matching model for microtubule segmentation. Unlike conventional U-Net variants that predict masks in a single pass, MTFlow learns vector fields that iteratively transport noisy masks toward the ground truth, enabling interpretable, trajectory-based refinement. Our architecture combines a U-Net backbone with temporal embeddings, allowing the model to capture the dynamics of uncertainty resolution along filament boundaries. We trained and evaluated MTFlow on synthetic and real microtubule datasets and assessed its generalization capability on public biomedical datasets of curvilinear structures such as retinal blood vessels and nerves. MTFlow achieves competitive segmentation accuracy comparable to state-of-the-art models, offering a powerful and time-efficient tool for filamentous structure analysis with more precise annotations than manual or semi-automatic approaches.

Achraf Ait Laydi, Sidi Mohamed Sid'El Moctar, Yousef El Mourabit et al.

Accurate quantification of the geometry of curvilinear biological structures is essential for understanding cellular mechanics and disease-related morphological alterations. Microtubule curvature is a key descriptor of filament rigidity and mechanical perturbations. However, reliable curvature extraction from fluorescence microscopy images remains challenging due to noise, low contrast, and partial filament visibility. Existing approaches rely on segmentation pipelines with pre or post-processing, which are highly sensitive to segmentation errors and often fail under adverse imaging conditions. In this work, we propose MTCurv, a deep learning framework for direct, segmenta-tion-free regression of microtubule curvature maps from noisy microscopy images. Leveraging a synthetic dataset with pixel-wise curvature annotations, we reformulated curvature estimation as a regression problem and adapted an attention-based residual U-Net. To reduce hallucinations and enforce spatial coherence, we introduced a gradient-aware loss combining Mean Squared Error with a gradient consistency term. Beyond model and loss design, we evaluated commonly used regression and image quality metrics, revealing that many perceptual and blind metrics are poorly suited for curvature estimation. Correlation-based metrics, particularly Spearman correlation, emerged as more reliable indicators of curvature prediction quality. Experiments on two datasets of increasing difficulty demonstrated that MTCurv accurately recovers local microtubule curvatures, even in the presence of background fluorescence. Ablation studies highlighted the contribution of both residual encoding and attention-based decoding. Overall, this work provides a practical tool for filament curvature analysis and methodological insights for geometry-aware regression in biomedical imaging. Datasets and code are made available.

Achraf Ait Laydi, Louis Cueff, Mewen Crespo et al.

Segmenting cytoskeletal filaments in microscopy images is essential for understanding their cellular roles but remains challenging, especially in dense, complex networks and under noisy or low-contrast image conditions. While deep learning has advanced image segmentation, performance often degrades in these adverse scenarios. Additional challenges include the difficulty of obtaining accurate annotations and managing severe class imbalance. We proposed a novel noise-adaptive attention mechanism, extending the Squeeze-and-Excitation (SE) module, to dynamically adjust to varying noise levels. This Adaptive SE (ASE) mechanism is integrated into a U-Net decoder, with residual encoder blocks, forming a lightweight yet powerful model: ASE_Res_U-Net. We also developed a synthetic-dataset strategy and employed tailored loss functions and evaluation metrics to mitigate class imbalance and ensure fair assessment. ASE_Res_U-Net effectively segmented microtubules in both synthetic and real noisy images, outperforming its ablated variants and state-of-the-art curvilinear-structure segmentation methods. It achieved this while using fewer parameters, making it suitable for resource-constrained environments. Importantly, ASE_Res_U-Net generalised well to other curvilinear structures (blood vessels and nerves) under diverse imaging conditions. Availability and implementation: Original microtubule datasets (synthetic and real noisy images) are available on Zenodo (DOIs: 10.5281/zenodo.14696279 and 10.5281/zenodo.15852660). ASE_Res_UNet model will be shared upon publication.