Digital Staining with Knowledge Distillation: A Unified Framework for Unpaired and Paired-But-Misaligned Data

Staining is essential in cell imaging and medical diagnostics but poses significant challenges, including high cost, time consumption, labor intensity, and irreversible tissue alterations. Recent advances in deep learning have enabled digital staining through supervised model training. However, collecting large-scale, perfectly aligned pairs of stained and unstained images remains difficult. In this work, we propose a novel unsupervised deep learning framework for digital cell staining that reduces the need for extensive paired data using knowledge distillation. We explore two training schemes: (1) unpaired and (2) paired-but-misaligned settings. For the unpaired case, we introduce a two-stage pipeline, comprising light enhancement followed by colorization, as a teacher model. Subsequently, we obtain a student staining generator through knowledge distillation with hybrid non-reference losses. To leverage the pixel-wise information between adjacent sections, we further extend to the paired-but-misaligned setting, adding the Learning to Align module to utilize pixel-level information. Experiment results on our dataset demonstrate that our proposed unsupervised deep staining method can generate stained images with more accurate positions and shapes of the cell targets in both settings. Compared with competing methods, our method achieves improved results both qualitatively and quantitatively (e.g., NIQE and PSNR).We applied our digital staining method to the White Blood Cell (WBC) dataset, investigating its potential for medical applications.