Deep learning EPI-TIRF cross-modality enables background subtraction and axial super-resolution for widefield fluorescence microscopy

• URL: http://arxiv.org/abs/2511.06853v1
• Date: Mon, 10 Nov 2025 08:52:56 GMT
• Title: Deep learning EPI-TIRF cross-modality enables background subtraction and axial super-resolution for widefield fluorescence microscopy
• Authors: Qiushi Li, Celi Lou, Yanfang Cheng, Bilang Gong, Xinlin Chen, Hao Chen, Baowan Li, Jieli Wang, Yulin Wang, Sipeng Yang, Yunqing Tang, Luru Dai,
• Abstract summary: We developed ET2dNet, a deep learning-based EPI-TIRF cross-modality network that achieves TIRF-comparable background subtraction and axial super-resolution.<n>We also developed ET3dNet, a dedicated three-dimensional reconstruction network that produces artifact-reduced results.
• Score: 22.180941479385755
• License: http://arxiv.org/licenses/nonexclusive-distrib/1.0/
• Abstract: The resolving ability of wide-field fluorescence microscopy is fundamentally limited by out-of-focus background owing to its low axial resolution, particularly for densely labeled biological samples. To address this, we developed ET2dNet, a deep learning-based EPI-TIRF cross-modality network that achieves TIRF-comparable background subtraction and axial super-resolution from a single wide-field image without requiring hardware modifications. The model employs a physics-informed hybrid architecture, synergizing supervised learning with registered EPI-TIRF image pairs and self-supervised physical modeling via convolution with the point spread function. This framework ensures exceptional generalization across microscope objectives, enabling few-shot adaptation to new imaging setups. Rigorous validation on cellular and tissue samples confirms ET2dNet's superiority in background suppression and axial resolution enhancement, while maintaining compatibility with deconvolution techniques for lateral resolution improvement. Furthermore, by extending this paradigm through knowledge distillation, we developed ET3dNet, a dedicated three-dimensional reconstruction network that produces artifact-reduced volumetric results. ET3dNet effectively removes out-of-focus background signals even when the input image stack lacks the source of background. This framework makes axial super-resolution imaging more accessible by providing an easy-to-deploy algorithm that avoids additional hardware costs and complexity, showing great potential for live cell studies and clinical histopathology.

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