Studying microtubules (MTs) and their mechanical properties is central to understanding intracellular transport, cell division, and drug action. While important, experts still need to spend many hours manually segmenting these filamentous structures. The suitability of state-of-the-art methods for this task cannot be systematically assessed, as large-scale labeled datasets are missing. We address this gap by introducing the synthetic dataset SynthMT , produced by tuning a novel image generation pipeline on real-world interference reflection microscopy (IRM) frames of in vitro reconstituted MTs without requiring human annotations. In our benchmark, we evaluate nine fully automated methods for MT analysis in both zero- and Hyperparameter Optimization (HPO)-based few-shot settings. Across both settings, classical algorithms and current foundation models still struggle to achieve the accuracy required for biological downstream analysis on in vitro MT IRM images that humans perceive as visually simple. However, a notable exception is the recently introduced SAM3 model. After HPO on only ten random SynthMT images, its text-prompted version SAM3Text achieves near-perfect and in some cases super-human performance on unseen, real data. This indicates that fully automated MT segmentation has become feasible when method configuration is effectively guided by synthetic data. To enable progress, we publicly release the dataset, the generation pipeline, and the evaluation framework.
Koddenbrock et al. (2026) studied this question.