Information-hiding cameras: Optical concealment of object information into ordinary images.

We introduce an information-hiding camera integrated with an electronic decoder that is jointly optimized through deep learning. This system uses a diffractive optical processor, which transforms and hides input images into ordinary-looking patterns that deceive/mislead observers. This information-hiding transformation is valid for infinitely many combinations of secret messages, transformed into ordinary-looking output images through passive light-matter interactions within the diffractive processor. By processing these output patterns, an electronic decoder network accurately reconstructs the original information hidden within the deceptive output. We demonstrated our approach by designing information-hiding diffractive cameras operating under various lighting conditions and noise levels, showing their robustness. We further extended this framework to multispectral operation, allowing the concealment and decoding of multiple images at different wavelengths, performed simultaneously. The feasibility of our framework was also validated experimentally using terahertz radiation. This optical encoder-electronic decoder-based codesign provides a high speed and energy efficient information-hiding camera, offering a powerful solution for visual information security.

Broadband nonlinear modulation of incoherent light using a transparent optoelectronic neuron array.

Nonlinear optical processing of ambient natural light is highly desired for computational imaging and sensing. Strong optical nonlinear response under weak broadband incoherent light is essential for this purpose. By merging 2D transparent phototransistors (TPTs) with liquid crystal (LC) modulators, we create an optoelectronic neuron array that allows self-amplitude modulation of spatially incoherent light, achieving a large nonlinear contrast over a broad spectrum at orders-of-magnitude lower intensity than achievable in most optical nonlinear materials. We fabricated a 10,000-pixel array of optoelectronic neurons, and experimentally demonstrated an intelligent imaging system that instantly attenuates intense glares while retaining the weaker-intensity objects captured by a cellphone camera. This intelligent glare-reduction is important for various imaging applications, including autonomous driving, machine vision, and security cameras. The rapid nonlinear processing of incoherent broadband light might also find applications in optical computing, where nonlinear activation functions for ambient light conditions are highly sought.

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.

Universal linear intensity transformations using spatially incoherent diffractive processors.

Under spatially coherent light, a diffractive optical network composed of structured surfaces can be designed to perform any arbitrary complex-valued linear transformation between its input and output fields-of-view (FOVs) if the total number (N) of optimizable phase-only diffractive features is ≥~2NiNo, where Ni and No refer to the number of useful pixels at the input and the output FOVs, respectively. Here we report the design of a spatially incoherent diffractive optical processor that can approximate any arbitrary linear transformation in time-averaged intensity between its input and output FOVs. Under spatially incoherent monochromatic light, the spatially varying intensity point spread function (H) of a diffractive network, corresponding to a given, arbitrarily-selected linear intensity transformation, can be written as H(m, n; m', n') = |h(m, n; m', n')|2, where h is the spatially coherent point spread function of the same diffractive network, and (m, n) and (m', n') define the coordinates of the output and input FOVs, respectively. Using numerical simulations and deep learning, supervised through examples of input-output profiles, we demonstrate that a spatially incoherent diffractive network can be trained to all-optically perform any arbitrary linear intensity transformation between its input and output if N ≥ ~2NiNo. We also report the design of spatially incoherent diffractive networks for linear processing of intensity information at multiple illumination wavelengths, operating simultaneously. Finally, we numerically demonstrate a diffractive network design that performs all-optical classification of handwritten digits under spatially incoherent illumination, achieving a test accuracy of >95%. Spatially incoherent diffractive networks will be broadly useful for designing all-optical visual processors that can work under natural light.

Rapid and stain-free quantification of viral plaque via lens-free holography and deep learning.

A plaque assay-the gold-standard method for measuring the concentration of replication-competent lytic virions-requires staining and usually more than 48 h of runtime. Here we show that lens-free holographic imaging and deep learning can be combined to expedite and automate the assay. The compact imaging device captures phase information label-free at a rate of approximately 0.32 gigapixels per hour per well, covers an area of about 30 × 30 mm2 and a 10-fold larger dynamic range of virus concentration than standard assays, and quantifies the infected area and the number of plaque-forming units. For the vesicular stomatitis virus, the automated plaque assay detected the first cell-lysing events caused by viral replication as early as 5 h after incubation, and in less than 20 h it detected plaque-forming units at rates higher than 90% at 100% specificity. Furthermore, it reduced the incubation time of the herpes simplex virus type 1 by about 48 h and that of the encephalomyocarditis virus by about 20 h. The stain-free assay should be amenable for use in virology research, vaccine development and clinical diagnosis.

Data-Class-Specific All-Optical Transformations and Encryption.

Diffractive optical networks provide rich opportunities for visual computing tasks. Here, data-class-specific transformations that are all-optically performed between the input and output fields-of-view (FOVs) of a diffractive network are presented. The visual information of the objects is encoded into the amplitude (A), phase (P), or intensity (I) of the optical field at the input, which is all-optically processed by a data-class-specific diffractive network. At the output, an image sensor-array directly measures the transformed patterns, all-optically encrypted using the transformation matrices preassigned to different data classes, i.e., a separate matrix for each data class. The original input images can be recovered by applying the correct decryption key (the inverse transformation) corresponding to the matching data class, while applying any other key will lead to loss of information. All-optical class-specific transformations covering A → A, I → I, and P → I transformations using various image datasets are numerically demonstrated. The feasibility of this framework is also experimentally validated by fabricating class-specific I → I transformation diffractive networks and is successfully tested at different parts of the electromagnetic spectrum, i.e., 1550 nm and 0.75 mm wavelengths. Data-class-specific all-optical transformations provide a fast and energy-efficient method for image and data encryption, enhancing data security and privacy.

Unidirectional imaging using deep learning-designed materials.

A unidirectional imager would only permit image formation along one direction, from an input field-of-view (FOV) A to an output FOV B, and in the reverse path, B â†’ A, the image formation would be blocked. We report the first demonstration of unidirectional imagers, presenting polarization-insensitive and broadband unidirectional imaging based on successive diffractive layers that are linear and isotropic. After their deep learning-based training, the resulting diffractive layers are fabricated to form a unidirectional imager. Although trained using monochromatic illumination, the diffractive unidirectional imager maintains its functionality over a large spectral band and works under broadband illumination. We experimentally validated this unidirectional imager using terahertz radiation, well matching our numerical results. We also created a wavelength-selective unidirectional imager, where two unidirectional imaging operations, in reverse directions, are multiplexed through different illumination wavelengths. Diffractive unidirectional imaging using structured materials will have numerous applications in, e.g., security, defense, telecommunications, and privacy protection.

Deep learning-enabled virtual histological staining of biological samples.

Histological staining is the gold standard for tissue examination in clinical pathology and life-science research, which visualizes the tissue and cellular structures using chromatic dyes or fluorescence labels to aid the microscopic assessment of tissue. However, the current histological staining workflow requires tedious sample preparation steps, specialized laboratory infrastructure, and trained histotechnologists, making it expensive, time-consuming, and not accessible in resource-limited settings. Deep learning techniques created new opportunities to revolutionize staining methods by digitally generating histological stains using trained neural networks, providing rapid, cost-effective, and accurate alternatives to standard chemical staining methods. These techniques, broadly referred to as virtual staining, were extensively explored by multiple research groups and demonstrated to be successful in generating various types of histological stains from label-free microscopic images of unstained samples; similar approaches were also used for transforming images of an already stained tissue sample into another type of stain, performing virtual stain-to-stain transformations. In this Review, we provide a comprehensive overview of the recent research advances in deep learning-enabled virtual histological staining techniques. The basic concepts and the typical workflow of virtual staining are introduced, followed by a discussion of representative works and their technical innovations. We also share our perspectives on the future of this emerging field, aiming to inspire readers from diverse scientific fields to further expand the scope of deep learning-enabled virtual histological staining techniques and their applications.

Roadmap on Deep Learning for Microscopy.

Through digital imaging, microscopy has evolved from primarily being a means for visual observation of life at the micro- and nano-scale, to a quantitative tool with ever-increasing resolution and throughput. Artificial intelligence, deep neural networks, and machine learning are all niche terms describing computational methods that have gained a pivotal role in microscopy-based research over the past decade. This Roadmap is written collectively by prominent researchers and encompasses selected aspects of how machine learning is applied to microscopy image data, with the aim of gaining scientific knowledge by improved image quality, automated detection, segmentation, classification and tracking of objects, and efficient merging of information from multiple imaging modalities. We aim to give the reader an overview of the key developments and an understanding of possibilities and limitations of machine learning for microscopy. It will be of interest to a wide cross-disciplinary audience in the physical sciences and life sciences.

All-optical image classification through unknown random diffusers using a single-pixel diffractive network.

Classification of an object behind a random and unknown scattering medium sets a challenging task for computational imaging and machine vision fields. Recent deep learning-based approaches demonstrated the classification of objects using diffuser-distorted patterns collected by an image sensor. These methods demand relatively large-scale computing using deep neural networks running on digital computers. Here, we present an all-optical processor to directly classify unknown objects through unknown, random phase diffusers using broadband illumination detected with a single pixel. A set of transmissive diffractive layers, optimized using deep learning, forms a physical network that all-optically maps the spatial information of an input object behind a random diffuser into the power spectrum of the output light detected through a single pixel at the output plane of the diffractive network. We numerically demonstrated the accuracy of this framework using broadband radiation to classify unknown handwritten digits through random new diffusers, never used during the training phase, and achieved a blind testing accuracy of 87.74 ± 1.12%. We also experimentally validated our single-pixel broadband diffractive network by classifying handwritten digits "0" and "1" through a random diffuser using terahertz waves and a 3D-printed diffractive network. This single-pixel all-optical object classification system through random diffusers is based on passive diffractive layers that process broadband input light and can operate at any part of the electromagnetic spectrum by simply scaling the diffractive features proportional to the wavelength range of interest. These results have various potential applications in, e.g., biomedical imaging, security, robotics, and autonomous driving.

Label-Free Virtual HER2 Immunohistochemical Staining of Breast Tissue using Deep Learning.

The immunohistochemical (IHC) staining of the human epidermal growth factor receptor 2 (HER2) biomarker is widely practiced in breast tissue analysis, preclinical studies, and diagnostic decisions, guiding cancer treatment and investigation of pathogenesis. HER2 staining demands laborious tissue treatment and chemical processing performed by a histotechnologist, which typically takes one day to prepare in a laboratory, increasing analysis time and associated costs. Here, we describe a deep learning-based virtual HER2 IHC staining method using a conditional generative adversarial network that is trained to rapidly transform autofluorescence microscopic images of unlabeled/label-free breast tissue sections into bright-field equivalent microscopic images, matching the standard HER2 IHC staining that is chemically performed on the same tissue sections. The efficacy of this virtual HER2 staining framework was demonstrated by quantitative analysis, in which three board-certified breast pathologists blindly graded the HER2 scores of virtually stained and immunohistochemically stained HER2 whole slide images (WSIs) to reveal that the HER2 scores determined by inspecting virtual IHC images are as accurate as their immunohistochemically stained counterparts. A second quantitative blinded study performed by the same diagnosticians further revealed that the virtually stained HER2 images exhibit a comparable staining quality in the level of nuclear detail, membrane clearness, and absence of staining artifacts with respect to their immunohistochemically stained counterparts. This virtual HER2 staining framework bypasses the costly, laborious, and time-consuming IHC staining procedures in laboratory and can be extended to other types of biomarkers to accelerate the IHC tissue staining used in life sciences and biomedical workflow.

Biopsy-free in vivo virtual histology of skin using deep learning.

An invasive biopsy followed by histological staining is the benchmark for pathological diagnosis of skin tumors. The process is cumbersome and time-consuming, often leading to unnecessary biopsies and scars. Emerging noninvasive optical technologies such as reflectance confocal microscopy (RCM) can provide label-free, cellular-level resolution, in vivo images of skin without performing a biopsy. Although RCM is a useful diagnostic tool, it requires specialized training because the acquired images are grayscale, lack nuclear features, and are difficult to correlate with tissue pathology. Here, we present a deep learning-based framework that uses a convolutional neural network to rapidly transform in vivo RCM images of unstained skin into virtually-stained hematoxylin and eosin-like images with microscopic resolution, enabling visualization of the epidermis, dermal-epidermal junction, and superficial dermis layers. The network was trained under an adversarial learning scheme, which takes ex vivo RCM images of excised unstained/label-free tissue as inputs and uses the microscopic images of the same tissue labeled with acetic acid nuclear contrast staining as the ground truth. We show that this trained neural network can be used to rapidly perform virtual histology of in vivo, label-free RCM images of normal skin structure, basal cell carcinoma, and melanocytic nevi with pigmented melanocytes, demonstrating similar histological features to traditional histology from the same excised tissue. This application of deep learning-based virtual staining to noninvasive imaging technologies may permit more rapid diagnoses of malignant skin neoplasms and reduce invasive skin biopsies.

Calcium pyrophosphate crystal size and characteristics.

OBJECTIVE: To describe the characteristics of calcium pyrophosphate (CPP) crystal size and morphology under compensated polarized light microscopy (CPLM). Secondarily, to describe CPP crystals seen only with digital enhancement of CPLM images, confirmed with advanced imaging techniques. METHODS: Clinical lab-identified CPP-positive synovial fluid samples were collected from 16 joint aspirates. Four raters used a standardized protocol to describe crystal shape, birefringence strength and color. A crystal expert confirmed CPLM-visualized crystal identification. For crystal measurement, a high-pass linear light filter was used to enhance resolution and line discrimination of digital images. This process identified additional enhanced crystals not seen by raters under CPLM. Single-shot computational polarized light microscopy (SCPLM) provided further confirmation of the enhanced crystals' presence. RESULTS: Of 932 suspected crystals identified by CPLM, 569 met our inclusion criteria, and 293 (51%) were confirmed as CPP crystals. Of 175 unique confirmed crystals, 118 (67%) were rods (median area 3.6 µm2 [range, 1.0-22.9 µm2]), and 57 (33%) were rhomboids (median area 4.8 µm2 [range, 0.9-16.7 µm2]). Crystals visualized only after digital image enhancement were smaller and less birefringent than CPLM-identified crystals. CONCLUSIONS: CPP crystals that are smaller and weakly birefringent are more difficult to identify. There is likely a population of smaller, less birefringent CPP crystals that routinely goes undetected by CPLM. Describing the characteristics of poorly visible crystals may be of use for future development of novel crystal identification methods.

Early detection and classification of live bacteria using time-lapse coherent imaging and deep learning.

Early identification of pathogenic bacteria in food, water, and bodily fluids is very important and yet challenging, owing to sample complexities and large sample volumes that need to be rapidly screened. Existing screening methods based on plate counting or molecular analysis present various tradeoffs with regard to the detection time, accuracy/sensitivity, cost, and sample preparation complexity. Here, we present a computational live bacteria detection system that periodically captures coherent microscopy images of bacterial growth inside a 60-mm-diameter agar plate and analyses these time-lapsed holograms using deep neural networks for the rapid detection of bacterial growth and the classification of the corresponding species. The performance of our system was demonstrated by the rapid detection of Escherichia coli and total coliform bacteria (i.e., Klebsiella aerogenes and Klebsiella pneumoniae subsp. pneumoniae) in water samples, shortening the detection time by >12 h compared to the Environmental Protection Agency (EPA)-approved methods. Using the preincubation of samples in growth media, our system achieved a limit of detection (LOD) of ~1 colony forming unit (CFU)/L in ≤9 h of total test time. This platform is highly cost-effective (~$0.6/test) and has high-throughput with a scanning speed of 24 cm2/min over the entire plate surface, making it highly suitable for integration with the existing methods currently used for bacteria detection on agar plates. Powered by deep learning, this automated and cost-effective live bacteria detection platform can be transformative for a wide range of applications in microbiology by significantly reducing the detection time and automating the identification of colonies without labelling or the need for an expert.

Deep Learning-Based Holographic Polarization Microscopy.

Polarized light microscopy provides high contrast to birefringent specimen and is widely used as a diagnostic tool in pathology. However, polarization microscopy systems typically operate by analyzing images collected from two or more light paths in different states of polarization, which lead to relatively complex optical designs, high system costs, or experienced technicians being required. Here, we present a deep learning-based holographic polarization microscope that is capable of obtaining quantitative birefringence retardance and orientation information of specimen from a phase-recovered hologram, while only requiring the addition of one polarizer/analyzer pair to an inline lensfree holographic imaging system. Using a deep neural network, the reconstructed holographic images from a single state of polarization can be transformed into images equivalent to those captured using a single-shot computational polarized light microscope (SCPLM). Our analysis shows that a trained deep neural network can extract the birefringence information using both the sample specific morphological features as well as the holographic amplitude and phase distribution. To demonstrate the efficacy of this method, we tested it by imaging various birefringent samples including, for example, monosodium urate and triamcinolone acetonide crystals. Our method achieves similar results to SCPLM both qualitatively and quantitatively, and due to its simpler optical design and significantly larger field-of-view this method has the potential to expand the access to polarization microscopy and its use for medical diagnosis in resource limited settings.

Pathological crystal imaging with single-shot computational polarized light microscopy.

Pathological crystal identification is routinely practiced in rheumatology for diagnosing arthritis disease such as gout, and relies on polarized light microscopy as the gold standard method used by medical professionals. Here, we present a single-shot computational polarized light microscopy method that reconstructs the transmittance, retardance and slow-axis orientation of a birefringent sample using a single image captured with a pixelated-polarizer camera. This method is fast, simple-to-operate and compatible with all the existing standard microscopes without extensive or costly modifications. We demonstrated the success of our method by imaging three different types of crystals found in synovial fluid and reconstructed the birefringence information of these samples using a single image, without being affected by the orientation of individual crystals within the sample field-of-view. We believe this technique will provide improved sensitivity, specificity and speed, all at low cost, for clinical diagnosis of crystals found in synovial fluid and other bodily fluids.

Computational cytometer based on magnetically modulated coherent imaging and deep learning.

Detecting rare cells within blood has numerous applications in disease diagnostics. Existing rare cell detection techniques are typically hindered by their high cost and low throughput. Here, we present a computational cytometer based on magnetically modulated lensless speckle imaging, which introduces oscillatory motion to the magnetic-bead-conjugated rare cells of interest through a periodic magnetic force and uses lensless time-resolved holographic speckle imaging to rapidly detect the target cells in three dimensions (3D). In addition to using cell-specific antibodies to magnetically label target cells, detection specificity is further enhanced through a deep-learning-based classifier that is based on a densely connected pseudo-3D convolutional neural network (P3D CNN), which automatically detects rare cells of interest based on their spatio-temporal features under a controlled magnetic force. To demonstrate the performance of this technique, we built a high-throughput, compact and cost-effective prototype for detecting MCF7 cancer cells spiked in whole blood samples. Through serial dilution experiments, we quantified the limit of detection (LoD) as 10 cells per millilitre of whole blood, which could be further improved through multiplexing parallel imaging channels within the same instrument. This compact, cost-effective and high-throughput computational cytometer can potentially be used for rare cell detection and quantification in bodily fluids for a variety of biomedical applications.

Motility-based label-free detection of parasites in bodily fluids using holographic speckle analysis and deep learning.

Parasitic infections constitute a major global public health issue. Existing screening methods that are based on manual microscopic examination often struggle to provide sufficient volumetric throughput and sensitivity to facilitate early diagnosis. Here, we demonstrate a motility-based label-free computational imaging platform to rapidly detect motile parasites in optically dense bodily fluids by utilizing the locomotion of the parasites as a specific biomarker and endogenous contrast mechanism. Based on this principle, a cost-effective and mobile instrument, which rapidly screens ~3.2 mL of fluid sample in three dimensions, was built to automatically detect and count motile microorganisms using their holographic time-lapse speckle patterns. We demonstrate the capabilities of our platform by detecting trypanosomes, which are motile protozoan parasites, with various species that cause deadly diseases affecting millions of people worldwide. Using a holographic speckle analysis algorithm combined with deep learning-based classification, we demonstrate sensitive and label-free detection of trypanosomes within spiked whole blood and artificial cerebrospinal fluid (CSF) samples, achieving a limit of detection of ten trypanosomes per mL of whole blood (~five-fold better than the current state-of-the-art parasitological method) and three trypanosomes per mL of CSF. We further demonstrate that this platform can be applied to detect other motile parasites by imaging Trichomonas vaginalis, the causative agent of trichomoniasis, which affects 275 million people worldwide. With its cost-effective, portable design and rapid screening time, this unique platform has the potential to be applied for sensitive and timely diagnosis of neglected tropical diseases caused by motile parasites and other parasitic infections in resource-limited regions.