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Deep learning enables cross-modality super-resolution in fluorescence microscopy


We present deep-learning-enabled super-resolution across different fluorescence microscopy modalities. This data-driven approach does not require numerical modeling of the imaging process or the estimation of a point-spread-function, and is based on training a generative adversarial network (GAN) to transform diffraction-limited input images into super-resolved ones. Using this framework, we improve the resolution of wide-field images acquired with low-numerical-aperture objectives, matching the resolution that is acquired using high-numerical-aperture objectives. We also demonstrate cross-modality super-resolution, transforming confocal microscopy images to match the resolution acquired with a stimulated emission depletion (STED) microscope. We further demonstrate that total internal reflection fluorescence (TIRF) microscopy images of subcellular structures within cells and tissues can be transformed to match the results obtained with a TIRF-based structured illumination microscope. The deep network rapidly outputs these super-resolved images, without any iterations or parameter search, and could serve to democratize super-resolution imaging.

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Fig. 1: Deep-learning-based super-resolved images of bovine pulmonary artery endothelial cells (BPAECs).
Fig. 2: Comparison of deep learning results against Lucy–Richardson (LR) and non-negative least square (NNLS) image deconvolution algorithms.
Fig. 3: Image resolution improvement beyond the diffraction limit: from confocal microscopy to STED.
Fig. 4: PSF characterization, before and after the network, and its comparison to STED.
Fig. 5: Deep-learning enabled cross-modality image transformation from confocal to STED.
Fig. 6: Deep-learning enabled cross-modality image transformation from TIRF to TIRF-SIM.

Data availability

We declare that all the data supporting the findings of this work are available within the manuscript and Supplementary Information files. Raw images can be requested from the corresponding author. Deep learning models reported in this work used standard libraries and scripts that are publicly available in TensorFlow. The instruction manual for our Fiji/ImageJ plugin and trained models (available online as Supplementary Software 1–7) is provided as a Supplementary Protocol.


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The Ozcan Research Group at UCLA acknowledges the support of NSF Engineering Research Center (ERC, PATHS-UP), the Army Research Office (ARO; W911NF-13-1-0419 and W911NF-13-1-0197), the ARO Life Sciences Division, the National Science Foundation (NSF) CBET Division Biophotonics Program, the NSF Emerging Frontiers in Research and Innovation (EFRI) Award, the NSF INSPIRE Award, NSF Partnerships for Innovation: Building Innovation Capacity (PFI:BIC) Program, the National Institutes of Health (NIH, R21EB023115), the Howard Hughes Medical Institute (HHMI), Vodafone Americas Foundation, the Mary Kay Foundation, and Steven & Alexandra Cohen Foundation. Yair Rivenson is partially supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No H2020-MSCA-IF-2014-659595 (MCMQCT). Confocal and STED laser scanning microscopy was performed at the California NanoSystems Institute (CNSI) Advanced Light Microscopy/Spectroscopy Shared Resource Facility at UCLA. We also thank the Advanced Imaging Center (AIC) at Janelia Research Campus for access to their TIRF-SIM microscope. The AIC is jointly supported by the Howard Hughes Medical Institute and the Gordon and Betty Moore Foundation. Finally, we thank H. Chang (Purdue University, West Lafayette, IN, USA) for sharing the CLC-mEmerald fly strain.

Author information




H.W., Y.R., and A.O. conceived the research. H.W., Y.R., L.B., and C.K. contributed to the experiments. H.W., Y.J., Z.W., and H.G. processed the data. H.W. and Y.J. prepared the figures. H.W., Y.R., and A.O. prepared the manuscript, and all the authors contributed to the manuscript. H.W., Y.R., and R.G. developed the Fiji/ImageJ plugin. A.O. supervised the research.

Corresponding author

Correspondence to Aydogan Ozcan.

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Competing interests

A.O., Y.R., and H.W. have a pending patent application on the contents of the presented results.

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Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary text and figures

Supplementary Notes 1–10, Supplementary Figures 1–14 and Supplementary Table 1

Reporting Summary

Supplementary Protocol

SISR-Fluorescent Fiji/ImageJ plugin: User Manual

Supplementary Video 1

Deep-learning enabled cross-modality image transformation from TIRF to TIRF-SIM. A video of deep-learning-enabled cross-modality image transformation from TIRF to TIRF-SIM, corresponding to a gene-edited SUM159 cell expressing AP2-eGFP, revealing the temporal dynamics of endocytic protein structures within the cell. The highlighted frames in this video correspond to subpanels of Fig. 6 (main text). Experiments were repeated with >1,000 images/frames, achieving similar results.

Supplementary Software 1

SISR-Fluorescent Fiji/ImageJ plugin

Supplementary Software 2

Pre-trained model: wide-field DAPI

Supplementary Software 3

Pre-trained model: wide-field FITC

Supplementary Software 4

Pre-trained model: wide-field TxRed

Supplementary Software 5

Pre-trained model: confocal STED (nanobeads)

Supplementary Software 6

Pre-trained model: confocal STED (nuclei)

Supplementary Software 7

Pre-trained model: TIRF-SIM

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Wang, H., Rivenson, Y., Jin, Y. et al. Deep learning enables cross-modality super-resolution in fluorescence microscopy. Nat Methods 16, 103–110 (2019).

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