Artificial intelligence-enabled quantitative phase imaging methods for life sciences.

Quantitative phase imaging, integrated with artificial intelligence, allows for the rapid and label-free investigation of the physiology and pathology of biological systems. This review presents the principles of various two-dimensional and three-dimensional label-free phase imaging techniques that exploit refractive index as an intrinsic optical imaging contrast. In particular, we discuss artificial intelligence-based analysis methodologies for biomedical studies including image enhancement, segmentation of cellular or subcellular structures, classification of types of biological samples and image translation to furnish subcellular and histochemical information from label-free phase images. We also discuss the advantages and challenges of artificial intelligence-enabled quantitative phase imaging analyses, summarize recent notable applications in the life sciences, and cover the potential of this field for basic and industrial research in the life sciences.

Improved algorithm of non-line-of-sight imaging based on the Bayesian statistics.

The recovery of obscured objects is an important goal in imaging that has been approached by exploiting coherence properties, ballistic photons, and penetrating wavelengths. In this paper, a robust reconstruction non-line-of-sight (NLOS) algorithm was proposed based on the Bayesian statistics, using the temporal, spatial, and intensity information of each signal. Compared with conventional back-projection methods, this algorithm is able to handle random errors in data and to image occluded objects with a higher quality. An adjustable compensation mechanism in our method is effective in dealing with the diversity of objects with regard to reflective characteristics. Our algorithm was demonstrated to be efficient with both simulated and experimental data. In addition, we discuss the advantages over existing methods and further improvement of the proposed algorithm.

Virtual histological staining of unlabeled autopsy tissue.

Traditional histochemical staining of post-mortem samples often confronts inferior staining quality due to autolysis caused by delayed fixation of cadaver tissue, and such chemical staining procedures covering large tissue areas demand substantial labor, cost and time. Here, we demonstrate virtual staining of autopsy tissue using a trained neural network to rapidly transform autofluorescence images of label-free autopsy tissue sections into brightfield equivalent images, matching hematoxylin and eosin (H&E) stained versions of the same samples. The trained model can effectively accentuate nuclear, cytoplasmic and extracellular features in new autopsy tissue samples that experienced severe autolysis, such as COVID-19 samples never seen before, where the traditional histochemical staining fails to provide consistent staining quality. This virtual autopsy staining technique provides a rapid and resource-efficient solution to generate artifact-free H&E stains despite severe autolysis and cell death, also reducing labor, cost and infrastructure requirements associated with the standard histochemical staining.

Light-field tomographic fluorescence lifetime imaging microscopy.

Fluorescence lifetime imaging microscopy (FLIM) is a powerful imaging technique that enables the visualization of biological samples at the molecular level by measuring the fluorescence decay rate of fluorescent probes. This provides critical information about molecular interactions, environmental changes, and localization within biological systems. However, creating high-resolution lifetime maps using conventional FLIM systems can be challenging, as it often requires extensive scanning that can significantly lengthen acquisition times. This issue is further compounded in three-dimensional (3D) imaging because it demands additional scanning along the depth axis. To tackle this challenge, we developed a novel computational imaging technique called light field tomographic FLIM (LIFT-FLIM). Our approach allows for the acquisition of volumetric fluorescence lifetime images in a highly data-efficient manner, significantly reducing the number of scanning steps required compared to conventional point-scanning or line-scanning FLIM imagers. Moreover, LIFT-FLIM enables the measurement of high-dimensional data using low-dimensional detectors, which are typically low-cost and feature a higher temporal bandwidth. We demonstrated LIFT-FLIM using a linear single-photon avalanche diode array on various biological systems, showcasing unparalleled single-photon detection sensitivity. Additionally, we expanded the functionality of our method to spectral FLIM and demonstrated its application in high-content multiplexed imaging of lung organoids. LIFT-FLIM has the potential to open up new avenues in both basic and translational biomedical research.

Roadmap on Label-Free Super-Resolution Imaging.

Label-free super-resolution (LFSR) imaging relies on light-scattering processes in nanoscale objects without a need for fluorescent (FL) staining required in super-resolved FL microscopy. The objectives of this Roadmap are to present a comprehensive vision of the developments, the state-of-the-art in this field, and to discuss the resolution boundaries and hurdles which need to be overcome to break the classical diffraction limit of the LFSR imaging. The scope of this Roadmap spans from the advanced interference detection techniques, where the diffraction-limited lateral resolution is combined with unsurpassed axial and temporal resolution, to techniques with true lateral super-resolution capability which are based on understanding resolution as an information science problem, on using novel structured illumination, near-field scanning, and nonlinear optics approaches, and on designing superlenses based on nanoplasmonics, metamaterials, transformation optics, and microsphere-assisted approaches. To this end, this Roadmap brings under the same umbrella researchers from the physics and biomedical optics communities in which such studies have often been developing separately. The ultimate intent of this paper is to create a vision for the current and future developments of LFSR imaging based on its physical mechanisms and to create a great opening for the series of articles in this field.

Fourier Imager Network (FIN): A deep neural network for hologram reconstruction with superior external generalization.

Deep learning-based image reconstruction methods have achieved remarkable success in phase recovery and holographic imaging. However, the generalization of their image reconstruction performance to new types of samples never seen by the network remains a challenge. Here we introduce a deep learning framework, termed Fourier Imager Network (FIN), that can perform end-to-end phase recovery and image reconstruction from raw holograms of new types of samples, exhibiting unprecedented success in external generalization. FIN architecture is based on spatial Fourier transform modules that process the spatial frequencies of its inputs using learnable filters and a global receptive field. Compared with existing convolutional deep neural networks used for hologram reconstruction, FIN exhibits superior generalization to new types of samples, while also being much faster in its image inference speed, completing the hologram reconstruction task in ~0.04 s per 1 mm2 of the sample area. We experimentally validated the performance of FIN by training it using human lung tissue samples and blindly testing it on human prostate, salivary gland tissue and Pap smear samples, proving its superior external generalization and image reconstruction speed. Beyond holographic microscopy and quantitative phase imaging, FIN and the underlying neural network architecture might open up various new opportunities to design broadly generalizable deep learning models in computational imaging and machine vision fields.

Recurrent neural network-based volumetric fluorescence microscopy.

Volumetric imaging of samples using fluorescence microscopy plays an important role in various fields including physical, medical and life sciences. Here we report a deep learning-based volumetric image inference framework that uses 2D images that are sparsely captured by a standard wide-field fluorescence microscope at arbitrary axial positions within the sample volume. Through a recurrent convolutional neural network, which we term as Recurrent-MZ, 2D fluorescence information from a few axial planes within the sample is explicitly incorporated to digitally reconstruct the sample volume over an extended depth-of-field. Using experiments on C. elegans and nanobead samples, Recurrent-MZ is demonstrated to significantly increase the depth-of-field of a 63×/1.4NA objective lens, also providing a 30-fold reduction in the number of axial scans required to image the same sample volume. We further illustrated the generalization of this recurrent network for 3D imaging by showing its resilience to varying imaging conditions, including e.g., different sequences of input images, covering various axial permutations and unknown axial positioning errors. We also demonstrated wide-field to confocal cross-modality image transformations using Recurrent-MZ framework and performed 3D image reconstruction of a sample using a few wide-field 2D fluorescence images as input, matching confocal microscopy images of the same sample volume. Recurrent-MZ demonstrates the first application of recurrent neural networks in microscopic image reconstruction and provides a flexible and rapid volumetric imaging framework, overcoming the limitations of current 3D scanning microscopy tools.

SNR-Adaptive OCT Angiography Enabled by Statistical Characterization of Intensity and Decorrelation With Multi-Variate Time Series Model.

In OCT angiography (OCTA), decorrelation computation has been widely used as a local motion index to identify dynamic flow from static tissues, but its dependence on SNR severely degrades the vascular visibility, particularly in low-SNR regions. To mathematically characterize the decorrelation-SNR dependence of OCT signals, we developed a multi-variate time series (MVTS) model. Based on the model, we derived a universal asymptotic linear relation of decorrelation to inverse SNR (iSNR), with the variance in static and noise regions determined by the average kernel size. Accordingly, with the population distribution of static and noise voxels being explicitly calculated in the iSNR and decorrelation (ID) space, a linear classifier is developed by removing static and noise voxels at all SNR, to generate a SNR-adaptive OCTA, termed as ID-OCTA. Then, flow phantom and human skin experiments were performed to validate the proposed ID-OCTA. Both qualitative and quantitative assessments demonstrated that the ID-OCTA offers a superior visibility of blood vessels, particularly in the deep layer. Finally, the implications of this work on both system design and hemodynamic quantification are further discussed.