Approaching human parity in the quality of automated organoid image segmentation
Authors: Chase Cartwright, Gongbo Guo, Sai Teja Pusuluri, Christopher N. Mayhew, Mark Hester, Horacio E. Castillo
Abstract
Organoids are complex, three dimensional, self-organizing cell cultures which manifest organ-like features and represent a powerful platform for studying human disease and developing treatment options. Organoid development is characterized by dynamic morphological and cellular organization, which mimic some aspects of organ development. To study these rapid changes over the course of organoid development, advanced imaging and analytical tools are critical to accurately monitor the trajectory of organoid growth and investigate disease processes. In this work, we focus on computer vision and machine learning techniques to automatically measure the size and shape of developing spheroids derived from pluripotent stem cells (iPSCs), which are typically the starting material for generating organoid cultures. To facilitate this task, we introduce a composite method that combines the Segment Anything Model (SAM), a general-purpose foundation model, with an existing domain-specific tool. This composite method is evaluated together with several existing tools by testing them on organoid image data and comparing with the results of manual image segmentation. We find that no single existing tool is able to segment the test images with sufficient accuracy across all test conditions, but the newly introduced composite method produces consistent and accurate results for all but a very small fraction of the most challenging images. Finally, we compare the accuracy of this method to the variability between manual segmentations by independent annotators (inter-observer variability) and find that by one measure it performs at the level of inter-observer variability and by others it performs very close to it.
Organoids are three-dimensional tissue models whose morphology provides important insights into tumor development, disease progression, and drug testing. Extracting these morphological features relies heavily on manual segmentation, which is time-consuming and labor-intensive. Furthermore, performing quantitative statistical analysis typically requires custom coding skills and a mathematical background, presenting a major barrier for experimental biologists. To address these challenges, we introduce MorphoOrgaAgent, a multi-agent framework that achieves zero-shot organoid segmentation, automated data analysis, and report generation based on natural language input. The framework consists mainly of three core components: a TaskUnderstandingAgent that identifies requested measurements and visualization types; a hybrid segmentation module that combines Cellpose-derived geometric prompts with text prompts to guide SAM3 for zero-shot organoid instance segmentation; and a ReportAgent that computes quantitative metrics and compiles them alongside generated visualizations into a structured report. We further introduce MorphoOrgaVQA, a benchmark designed for quantitative evaluation of agent systems in organoid morphology analysis. Experimental results demonstrate that MorphoOrgaAgent handles both explicit and descriptive user requests, produces measurements closely matching ground truth, and generates complete analysis reports without requiring manual programming. The complete source code and MorphoOrgaVQA benchmark are publicly available at https://github.com/peng-lab/MorphoOrgaAgent.
Hanyi Zhang, Maximilian Hoermann, Lion J. Gleiter +5
Accurate 3D plant organ segmentation is fundamental to automated phenotyping. Existing approaches rely on annotated training data or species-specific model configurations. We present an annotation-free pipeline for 3D plant organ segmentation, combining text-prompted SAM3 segmentation with semantic neural radiance fields (NeRFs). Given only multi-view RGB images and a list of class names, our zero-shot pipeline produces semantically labeled 3D point clouds without manual annotation, per-species fine-tuning, or domain-specific preprocessing. Multi-view NeRF fusion acts as effective implicit consensus mechanism that lifts imperfect per-frame masks into accurate 3D labels. On a controlled Begonia maculata testbed the SAM3 pipeline achieves 92.6% mIoU, reaching 95.9% of the oracle upper bound established with perfect ground-truth masks. The pipeline was further evaluated on a new dataset spanning ten diverse plant point clouds reaching an average 0.856 mIoU, with leaf and pot IoU above 0.91 and 0.90 for every species, respectively. These results demonstrate that annotation-free 3D plant organ segmentation is now feasible and approaching the range of supervised methods.
Foreground segmentation is the critical first step of every computational pathology pipeline, yet existing methods rely on hand-tuned heuristics or supervised models that overfit to narrow stain and scanner distributions, failing silently on specialised stains such as Jones silver or Elastica van Gieson. We propose a coarse-to-fine approach that recasts foreground segmentation as a visual perception task and leverages general-purpose vision-language models (VLMs) as zero-annotation oracles. Our key insight is that tissue-versus-background discrimination is a natural-image recognition problem, not a histopathological one, so VLMs trained on internet-scale corpora generalise where domain-specific models cannot. We introduce Leica-75, a benchmark of 75 renal transplant whole-slide images spanning three stain families. On Leica-75, our method achieves the highest segmentation quality on out-of-distribution stains (Dice 0.858 +/- 0.027 on Jones, 0.853 +/- 0.041 on EVG) with 7x lower cross-stain variance than the best supervised baseline, while remaining competitive on in-distribution H&E. Few-shot prompting with automatically curated exemplars (Auto-context) rescues hard cases on Stress-32 (n=32), a curated stress-test subset (Dice 0.470 to 0.819 for the 2B model). VLM-based annotation review matches human expert consensus (kappa=0.989 for blur detection; mean precision/recall grading accuracy 0.708 vs. human 0.646 for segmentation mask review). The resulting pseudo-labels are used to distil lightweight student models that are as performant as the teacher model while running for a fraction of the cost. Our framework provides a principled, scalable solution to a persistent infrastructure bottleneck in digital pathology.