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Inverse Protocol Prediction from Spheroid Microscopy Imaging via Morphology-Aware Structured Learning

Mittal, P.; Srivastava, A.; Chauhan, J.

2026-03-07 bioinformatics
10.64898/2026.03.04.709682 bioRxiv
Show abstract

We introduce Inverse Protocol Prediction (IPP), which is a task of inferring experimental culture conditions directly from a single bright-field spheroid image. We formulate IPP as a structured multi-label prediction problem and propose a protocol-aware learning framework that integrates morphology extraction, multimodal representation learning, and dependency-aware inference. Morphometric descriptors derived from automated spheroid segmentation are fused with deep visual embeddings via a morphometry vision fusion module. To capture biological and procedural dependencies, we develop a Hierarchical Multi-Task Transformer that conditions predictions across protocol attributes. The framework is trained with domainadversarial supervision and morphology-preserving augmentation to improve robustness to acquisition variability. Hybrid convolution attention encoders achieve the best performance, reaching 95.7% multi-attribute accuracy. We further evaluate cross dataset transfer and temporal morphology forecasting. Results demonstrate that structured, dependency-aware modeling enables reliable reconstruction of experimental protocols from imaging alone, supporting reproducibility auditing and protocol validation in 3D cell culture systems.

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