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Significance: Spatial frequency-domain imaging (SFDI) is a powerful technique for mapping tissue oxygen saturation over a wide field of view. However, current SFDI methods either require a sequence of several images with different illumination patterns or, in the case of single-snapshot optical properties (SSOP), introduce artifacts and sacrifice accuracy. Aim: We introduce OxyGAN, a data-driven, content-aware method to estimate tissue oxygenation directly from single structured-light images. Approach: OxyGAN is an end-to-end approach that uses supervised generative adversarial networks. Conventional SFDI is used to obtain ground truth tissue oxygenation maps for ex vivo human esophagi, in vivo hands and feet, and an in vivo pig colon sample under 659- and 851-nm sinusoidal illumination. We benchmark OxyGAN by comparing it with SSOP and a two-step hybrid technique that uses a previously developed deep learning model to predict optical properties followed by a physical model to calculate tissue oxygenation. Results: When tested on human feet, cross-validated OxyGAN maps tissue oxygenation with an accuracy of 96.5%. When applied to sample types not included in the training set, such as human hands and pig colon, OxyGAN achieves a 93% accuracy, demonstrating robustness to various tissue types. On average, OxyGAN outperforms SSOP and a hybrid model in estimating tissue oxygenation by 24.9% and 24.7%, respectively. Finally, we optimize OxyGAN inference so that oxygenation maps are computed ∼10 times faster than previous work, enabling video-rate, 25-Hz imaging. Conclusions: Due to its rapid acquisition and processing speed, OxyGAN has the potential to enable real-time, high-fidelity tissue oxygenation mapping that may be useful for many clinical applications.

Biomedical photoacoustic tomography, which can provide high resolution 3D soft tissue images based on the optical absorption, has advanced to the stage at which translation from the laboratory to clinical settings is becoming possible. The need for rapid image formation and the practical restrictions on data acquisition that arise from the constraints of a clinical workflow are presenting new image reconstruction challenges. There are many classical approaches to image reconstruction, but ameliorating the effects of incomplete or imperfect data through the incorporation of accurate priors is challenging and leads to slow algorithms. Recently, the application of Deep Learning, or deep neural networks, to this problem has received a great deal of attention. This paper reviews the literature on learned image reconstruction, summarising the current trends, and explains how these new approaches fit within, and to some extent have arisen from, a framework that encompasses classical reconstruction methods. In particular, it shows how these new techniques can be understood from a Bayesian perspective, providing useful insights. The paper also provides a concise tutorial demonstration of three prototypical approaches to learned image reconstruction. The code and data sets for these demonstrations are available to researchers. It is anticipated that it is in in vivo applications - where data may be sparse, fast imaging critical and priors difficult to construct by hand - that Deep Learning will have the most impact. With this in mind, the paper concludes with some indications of possible future research directions.

Fluorescence microscopy plays a vital role in understanding the subcellular structures of living cells. However, it requires considerable effort in sample preparation related to chemical fixation, staining, cost, and time. To reduce those factors, we present a virtual fluorescence staining method based on deep neural networks (VirFluoNet) to transform fluorescence images of molecular labels into other molecular fluorescence labels in the same field-of-view. To achieve this goal, we develop and train a conditional generative adversarial network (cGAN) to perform digital fluorescence imaging demonstrated on human osteosarcoma U2OS cell fluorescence images captured under Cell Painting staining protocol. A detailed comparative analysis is also conducted on the performance of the cGAN network between predicting fluorescence channels based on phase contrast or based on another fluorescence channel using human breast cancer MDA-MB-231 cell line as a test case. In addition, we implement a deep learning model to perform autofocusing on another human U2OS fluorescence dataset as a preprocessing step to defocus an out-focus channel in U2OS dataset. A quantitative index of image prediction error is introduced based on signal pixel-wise spatial and intensity differences with ground truth to evaluate the performance of prediction to high-complex and throughput fluorescence. This index provides a rational way to perform image segmentation on error signals and to understand the likelihood of mis-interpreting biology from the predicted image. In total, these findings contribute to the utility of deep learning image regression for fluorescence microscopy datasets of biological cells, balanced against savings of cost, time, and experimental effort. Furthermore, the approach introduced here holds promise for modeling the internal relationships between organelles and biomolecules within living cells., 20 pages, 9 figures