Abstract
Fluorescence microscopy-based image restoration has received widespread attention in the life sciences and has led to significant progress, benefiting from deep learning technology. However, most current task-specific methods have limited generalizability to different fluorescence microscopy-based image restoration problems. Here, we seek to improve generalizability and explore the potential of applying a pretrained foundation model to fluorescence microscopy-based image restoration. We provide a universal fluorescence microscopy-based image restoration (UniFMIR) model to address different restoration problems, and show that UniFMIR offers higher image restoration precision, better generalization and increased versatility. Demonstrations on five tasks and 14 datasets covering a wide range of microscopy imaging modalities and biological samples demonstrate that the pretrained UniFMIR can effectively transfer knowledge to a specific situation via fine-tuning, uncover clear nanoscale biomolecular structures and facilitate high-quality imaging. This work has the potential to inspire and trigger new research highlights for fluorescence microscopy-based image restoration.
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Data availability
All training and testing data involved in the experiments come from existing literature and can be downloaded from the corresponding links provided in Supplementary Table 2 or via Zenodo at https://doi.org/10.5281/zenodo.8401470 (ref. 55).
Code availability
The PyTorch code of our UniFMIR, together with trained models, as well as some example images for inference are publicly available at https://github.com/cxm12/UNiFMIR (https://doi.org/10.5281/zenodo.10117581)56. Furthermore, We also provide a live demo for UniFMIR at http://unifmir.fdudml.cn/. Users can also access the colab at https://colab.research.google.com/github/cxm12/UNiFMIR/blob/main/UniFMIR.ipynb or use the steps in our GitHub documentation to run the demo locally. This newly built interactive software platform facilitates users to freely and easily use the pretrained foundation model. It also makes it easy for us to continuously train the foundation model with new data and share it with the community. Finally, we shared all models on BioImage.IO at https://bioimage.io/#/. Data are available via Zenodo at https://doi.org/10.5281/zenodo.10577218, https://doi.org/10.5281/zenodo.10579778, https://doi.org/10.5281/zenodo.10579822, https://doi.org/10.5281/zenodo.10595428, https://doi.org/10.5281/zenodo.10595460, https://doi.org/10.5281/zenodo.8420081 and https://doi.org/10.5281/zenodo.8420100 (refs. 57,58,59,60,61,62,63). We used the Pycharm software for code development.
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Acknowledgements
We gratefully acknowledge support for this work provided by the National Natural Science Foundation of China (NSFC) (grant nos. U2001209 to B.Y. and 62372117 to W.T.) and the Natural Science Foundation of Shanghai (grant no. 21ZR1406600 to W.T.).
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B.Y. and W.T. supervised the research. C.M. and W.T. conceived of the technique. C.M. implemented the algorithm. C.M. and W.T. designed the validation experiments. C.M. trained the network and performed the validation experiments. R.H. implemented the interactive software platform and organized the codes and models. All authors had access to the study and wrote the paper.
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Extended data
Extended Data Fig. 1 Overall architecture of the UniFMIR.
The proposed UniFMIR approach is composed of three submodules: a multihead module, a Swin transformer-based feature enhancement module, and a multitail module. The numbers of parameters (M) and calculations (GFLOPs) required for the head, feature enhancement and tail modules for different tasks are marked below the structures of the respective modules. The input sizes and output sizes of training batches for different tasks are also marked below the images.
Extended Data Fig. 2 Network architecture of the Swin transformer-based feature enhancement module46.
The feature enhancement module consists of convolutional layers and a series of Swin transformer blocks (STB), each of which includes several Swin transformer layers (STL), a convolutional layer and a residual connection. The STL is composed of layer normalization operations, a multihead self-attention (MSA) mechanism and a multilayer perceptron (MLP). In the MSA mechanism, the input features are first divided into multiple small patches with a moving window operation, and then the self-attention in each patch is calculated to output features fout. The MLP is composed of two fully connected layers (FCs) and Gaussian-error linear unit (GELU) activation.
Extended Data Fig. 3 Generalization ability analysis of super-resolution on unseen modality of single-molecule localization microscopy data from the Shareloc platform52.
a, SR results obtained by the SOTA model (DeepSTORM54), the pretrained UniFMIR model without fine-tuning, Baseline (same network structure as UniFMIR trained from scratch), and our fine-tuned UniFMIR model. The GT dSTORM images of microtubules stained with Alexa 647 in U2OS cells incubated with nocodazole and the input synthesized LR images are also shown. The PSNR/NRMSE results of the SR outputs obtained on n = 16 synthetic inputs are shown on the right. b, SR results obtained on the real-world wide-field images. The NRMSE values are depicted on the residual images under different SR results and the raw input images. The PSNR/NRMSE results on n = 9 real-world inputs are shown on the right. Box-plot elements are defined as follows: center line (median); box limits (upper and lower quartiles); whiskers (1.5x interquartile range). The line plots show the pixel intensities along the dashed lines in the corresponding images. Scale bar: 6.5 μm.
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Supplementary Notes 1–5, Figs. 1–17 and Tables 1 and 2.
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Ma, C., Tan, W., He, R. et al. Pretraining a foundation model for generalizable fluorescence microscopy-based image restoration. Nat Methods (2024). https://doi.org/10.1038/s41592-024-02244-3
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DOI: https://doi.org/10.1038/s41592-024-02244-3