Significance: “Difference imaging,” which reconstructs target optical properties using measurements with and without target information, is often used in diffuse optical tomography (DOT) in vivo imaging. However, taking additional reference measurements is time consuming, and mismatches between the target medium and the reference medium can cause inaccurate reconstruction.

Aim: We aim to streamline the data acquisition and mitigate the mismatch problems in DOT difference imaging using a deep learning-based approach to generate data from target measurements only.

Approach: We train an artificial neural network to output data for difference imaging from target measurements only. The model is trained and validated on simulation data and tested with simulations, phantom experiments, and clinical data from 56 patients with breast lesions.

Results: The proposed method has comparable performance to the traditional approach using measurements without mismatch between the target side and the reference side, and it outperforms the traditional approach using measurements when there is a mismatch. It also improves the target-to-artifact ratio and lesion localization in patient data.

Conclusions: The proposed method can simplify the data acquisition procedure, mitigate mismatch problems, and improve reconstructed image quality in DOT difference imaging.