Search

Non-line-of-sight (NLOS) imaging enables the visualization of objects hidden from direct view, with applications in surveillance, remote sensing, and light detection and ranging. Here, we introduce a NLOS imaging technique termed ptychographic NLOS (pNLOS), which leverages coded ptychography for depth-resolved imaging of obscured objects. Our approach involves scanning a laser spot on a wall to illuminate the hidden objects in an obscured region. The reflected wavefields from these objects then travel back to the wall, get modulated by the wall's complex-valued profile, and the resulting diffraction patterns are captured by a camera. By modulating the object wavefields, the wall surface serves the role of the coded layer as in coded ptychography. As we scan the laser spot to different positions, the reflected object wavefields on the wall translate accordingly, with the shifts varying for objects at different depths. This translational diversity enables the acquisition of a set of modulated diffraction patterns referred to as a ptychogram. By processing the ptychogram, we recover both the objects at different depths and the modulation profile of the wall surface. Experimental results demonstrate high-resolution, high-fidelity imaging of hidden objects, showcasing the potential of pNLOS for depth-aware vision beyond the direct line of sight.

Synthetic aperture radar (SAR) utilizes an aircraft-carried antenna to emit electromagnetic pulses and detect the returning echoes. As the aircraft travels across a designated area, it synthesizes a large virtual aperture to improve image resolution. Inspired by SAR, we introduce synthetic aperture ptycho-endoscopy (SAPE) for micro-endoscopic imaging beyond the diffraction limit. SAPE operates by hand-holding a lensless fiber bundle tip to record coherent diffraction patterns from specimens. The fiber cores at the distal tip modulate the diffracted wavefield within a confined area, emulating the role of the 'airborne antenna' in SAR. The handheld operation introduces positional shifts to the tip, analogous to the aircraft's movement. These shifts facilitate the acquisition of a ptychogram and synthesize a large virtual aperture extending beyond the bundle's physical limit. We mitigate the influences of hand motion and fiber bending through a low-rank spatiotemporal decomposition of the bundle's modulation profile. Our tests demonstrate the ability to resolve a 548-nm linewidth on a resolution target. The achieved space-bandwidth product is ~1.1 million effective pixels, representing a 36-fold increase compared to that of the original fiber bundle. Furthermore, SAPE's refocusing capability enables imaging over an extended depth of field exceeding 2 cm. The aperture synthesizing process in SAPE surpasses the diffraction limit set by the probe's maximum collection angle, opening new opportunities for both fiber-based and distal-chip endoscopy in applications such as medical diagnostics and industrial inspection.

Fourier ptychography (FP) is an enabling imaging technique that produces high-resolution complex-valued images with extended field coverages. However, when FP images a phase object with any specific spatial frequency, the captured images contain only constant values, rendering the recovery of the corresponding linear phase ramp impossible. This challenge is not unique to FP but also affects other common microscopy techniques -- a rather counterintuitive outcome given their widespread use in phase imaging. The underlying issue originates from the non-uniform phase transfer characteristic inherent in microscope systems, which impedes the conversion of object wavefields into discernible intensity variations. To address this challenge, we present spatially-coded Fourier ptychography (scFP), a new method that synergizes FP with spatial-domain coded detection for true quantitative phase imaging. In scFP, a flexible and detachable coded thin film is attached atop the image sensor in a regular FP setup. The spatial modulation of this thin film ensures a uniform phase response across the entire synthetic bandwidth. It improves reconstruction quality and corrects refractive index underestimation issues prevalent in conventional FP and related tomographic implementations. The inclusion of the coded thin film further adds a new dimension of measurement diversity in the spatial domain. The development of scFP is expected to catalyse new research directions and applications for phase imaging, emphasizing the need for true quantitative accuracy with uniform frequency response.

Coded ptychography has emerged as a powerful technique for high-throughput, high-resolution lensless imaging. However, the trade-off between acquisition speed and image quality remains a significant challenge. To address this, we introduce a novel sparsity-regularized approach to coded ptychography that dramatically reduces the number of required measurements while maintaining high reconstruction quality. The reported approach, termed the ptychographic proximal total-variation (PPTV) solver, formulates the reconstruction task as a total variation regularized optimization problem. Unlike previous implementations that rely on specialized hardware or illumination schemes, PPTV integrates seamlessly into existing coded ptychography setups. Through comprehensive numerical simulations, we demonstrate that PPTV-driven coded ptychography can produce accurate reconstructions with as few as eight intensity measurements, a significant reduction compared to conventional methods. Convergence analysis confirms the robustness and stability of the PPTV algorithm. Experimental results from our optical prototype, featuring a disorder-engineered surface for wavefront modulation, validate PPTV's ability to achieve high-throughput, high-resolution imaging with a substantially reduced measurement burden. By enabling high-quality reconstructions from fewer measurements, PPTV paves the way for more compact, efficient, and cost-effective lensless microscopy systems on a chip, with potential applications in digital pathology, endoscopy, point-of-care diagnostics, and high-content screening., Comment: 14 pages, 7 figures

Polarimetric imaging provides valuable insights into the polarization state of light interacting with a sample. It can infer crucial birefringence properties of bio-specimens without using any labels, thereby facilitating the diagnosis of diseases such as cancer and osteoarthritis. In this study, we present a novel polarimetric coded ptychography (pol-CP) approach that enables high-resolution, high-throughput gigapixel birefringence imaging on a chip. Our platform deviates from traditional lens-based polarization systems by employing an integrated polarimetric coded sensor for lensless coherent diffraction imaging. Utilizing Jones calculus, we quantitatively determine the birefringence retardance and orientation information of bio-specimens from the recovered images. Our portable pol-CP prototype can resolve the 435-nm linewidth on the resolution target and the imaging field of view for a single acquisition is limited only by the detector size of 41^2. The prototype allows for the acquisition of gigapixel birefringence images with a 180-mm^2 field of view in ~3.5 minutes, a performance that rivals high-end whole slide scanner but a small fraction of the cost. To demonstrate its biomedical applications, we perform high-throughput imaging of malaria-infected blood smears, locating parasites using birefringence contrast. We also generate birefringence maps of label-free thyroid smears to identify thyroid follicles. Notably, the recovered birefringence maps emphasize the same regions as autofluorescence images, underscoring the potential for rapid on-site evaluation of label-free biopsies. Our approach provides a turnkey and portable solution for lensless polarimetric analysis on a chip, with promising applications in disease diagnosis, crystal screening, and label-free chemical imaging, particularly in resource-constrained environments.

Until recently, conventional biochemical staining had the undisputed status as well-established benchmark for most biomedical problems related to clinical diagnostics, fundamental research and biotechnology. Despite this role as gold-standard, staining protocols face several challenges, such as a need for extensive, manual processing of samples, substantial time delays, altered tissue homeostasis, limited choice of contrast agents for a given sample, 2D imaging instead of 3D tomography and many more. Label-free optical technologies, on the other hand, do not rely on exogenous and artificial markers, by exploiting intrinsic optical contrast mechanisms, where the specificity is typically less obvious to the human observer. Over the past few years, digital staining has emerged as a promising concept to use modern deep learning for the translation from optical contrast to established biochemical contrast of actual stainings. In this review article, we provide an in-depth analysis of the current state-of-the-art in this field, suggest methods of good practice, identify pitfalls and challenges and postulate promising advances towards potential future implementations and applications., Comment: Review article, 4 main Figures, 3 Tables, 2 supplementary figures

Blu-ray drive is an engineering masterpiece that integrates disc rotation, pickup head translation, and three lasers in a compact and portable format. Here we integrate a blood-coated image sensor with a modified Blu-ray drive for high-throughput cytometric analysis of various bio-specimens. In this device, samples are mounted on the rotating Blu-ray disc and illuminated by the built-in lasers from the pickup head. The resulting coherent diffraction patterns are then recorded by the blood-coated image sensor. The rich spatial features of the blood-cell monolayer help down-modulate the object information for sensor detection, thus forming a high-resolution computational bio-lens with a theoretically unlimited field of view. With the acquired data, we develop a lensless coherent diffraction imaging modality termed rotational ptychography for image reconstruction. We show that our device can resolve the 435 nm linewidth on the resolution target and has a field of view only limited by the size of the Blu-ray disc. To demonstrate its applications, we perform high-throughput urinalysis by locating disease-related calcium oxalate crystals over the entire microscope slide. We also quantify different types of cells on a blood smear with an acquisition speed of ~10,000 cells per second. For in vitro experiment, we monitor live bacterial cultures over the entire Petri dish with single-cell resolution. Using biological cells as a computational lens could enable new intriguing imaging devices for point-of-care diagnostics. Modifying a Blu-ray drive with the blood-coated sensor further allows the spread of high-throughput optical microscopy from well-equipped laboratories to citizen scientists worldwide.

We report the implementation of a fully on-chip, lensless microscopy technique termed optofluidic ptychography. This imaging modality complements the miniaturization provided by microfluidics and allows the integration of ptychographic microscopy into various lab-on-a-chip devices. In our prototype, we place a microfluidic channel on the top surface of a coverslip and coat the bottom surface with a scattering layer. The channel and the coated coverslip substrate are then placed on top of an image sensor for diffraction data acquisition. Similar to the operation of flow cytometer, the device utilizes microfluidic flow to deliver specimens across the channel. The diffracted light from the flowing objects is modulated by the scattering layer and recorded by the image sensor for ptychographic reconstruction, where high-resolution quantitative complex images are recovered from the diffraction measurements. By using an image sensor with a 1.85-micron pixel size, our device can resolve the 550 nm linewidth on the resolution target. We validate the device by imaging different types of biospecimens, including C. elegans, yeast cells, paramecium, and closterium sp. We also demonstrate high-resolution ptychographic reconstruction at a video framerate of 30 frames per second. The reported technique can address a wide range of biomedical needs and engenders new ptychographic imaging innovations in a flow cytometer configuration., Comment: 8 pages; 7 figures

Ptychography is an enabling coherent diffraction imaging technique for both fundamental and applied sciences. Its applications in optical microscopy, however, fall short for its low imaging throughput and limited resolution. Here, we report a resolution-enhanced parallel coded ptychography technique achieving the highest numerical aperture and an imaging throughput orders of magnitude greater than previous demonstrations. In this platform, we translate the samples across the disorder-engineered surfaces for lensless diffraction data acquisition. The engineered surface consists of chemically etched micron-level phase scatters and printed sub-wavelength intensity absorbers. It is designed to unlock an optical space with spatial extent (x, y) and frequency content (kx, ky) that is inaccessible using conventional lens-based optics. To achieve the best resolution performance, we also report a new coherent diffraction imaging model by considering both the spatial and angular responses of the pixel readouts. Our low-cost prototype can directly resolve 308-nm linewidth on the resolution target without aperture synthesizing. Gigapixel high-resolution microscopic images with a 240-mm^2 effective field of view can be acquired in 15 seconds. For demonstrations, we recover slow-varying 3D phase objects with many 2{\pi} wraps, including optical prism and convex lens. The low-frequency phase contents of these objects are challenging to obtain using other existing lensless techniques. For digital pathology applications, we perform accurate virtual staining by using the recovered phase as attention guidance in a deep neural network. Parallel optical processing using the reported technique enables novel optical instruments with inherent quantitative nature and metrological versatility., Comment: 17 pages; 7 figures

Traditional microbial detection methods often rely on the overall property of microbial cultures and cannot resolve individual growth event at high spatiotemporal resolution. As a result, they require bacteria to grow to confluence and then interpret the results. Here, we demonstrate the application of an integrated ptychographic sensor for lensless cytometric analysis of microbial cultures over a large scale and with high spatiotemporal resolution. The reported device can be placed within a regular incubator or used as a standalone incubating unit for long-term microbial monitoring. For longitudinal study where massive data are acquired at sequential time points, we report a new temporal-similarity constraint to increase the temporal resolution of ptychographic reconstruction by 7-fold. With this strategy, the reported device achieves a centimeter-scale field of view, a half-pitch spatial resolution of 488 nm, and a temporal resolution of 15-second intervals. For the first time, we report the direct observation of bacterial growth in a 15-second interval by tracking the phase wraps of the recovered images, with high phase sensitivity like that in interferometric measurements. We also characterize cell growth via longitudinal dry mass measurement and perform rapid bacterial detection at low concentrations. For drug-screening application, we demonstrate proof-of-concept antibiotic susceptibility testing and perform single-cell analysis of antibiotic-induced filamentation. The combination of high phase sensitivity, high spatiotemporal resolution, and large field of view is unique among existing microscopy techniques. As a quantitative and miniaturized platform, it can improve studies with microorganisms and other biospecimens at resource-limited settings., Comment: 18 pages, 6 figures

We report a new lensless microscopy configuration by integrating the concepts of transverse translational ptychography and defocus multi-height phase retrieval. In this approach, we place a tilted image sensor under the specimen for linearly-increasing phase modulation along one lateral direction. Similar to the operation of ptychography, we laterally translate the specimen and acquire the diffraction images for reconstruction. Since the axial distance between the specimen and the sensor varies at different lateral positions, laterally translating the specimen effectively introduces defocus multi-height measurements while eliminating axial scanning. Lateral translation further introduces sub-pixel shift for pixel super-resolution imaging and naturally expands the field of view for rapid whole slide imaging. We show that the equivalent height variation can be precisely estimated from the lateral shift of the specimen, thereby addressing the challenge of precise axial positioning in conventional multi-height phase retrieval. Using a sensor with a 1.67-micron pixel size, our low-cost and field-portable prototype can resolve 690-nm linewidth on the resolution target. We show that a whole slide image of a blood smear with a 120-mm^2 field of view can be acquired in 18 seconds. We also demonstrate accurate automatic white blood cell counting from the recovered image. The reported approach may provide a turnkey solution for addressing point-of-care- and telemedicine-related challenges.

Artificial Intelligence (AI)-powered pathology is a revolutionary step in the world of digital pathology and shows great promise to increase both diagnosis accuracy and efficiency. However, defocus and motion blur can obscure tissue or cell characteristics hence compromising AI algorithms'accuracy and robustness in analyzing the images. In this paper, we demonstrate a deep-learning-based approach that can alleviate the defocus and motion blur of a microscopic image and output a sharper and cleaner image with retrieved fine details without prior knowledge of the blur type, blur extent and pathological stain. In this approach, a deep learning classifier is first trained to identify the image blur type. Then, two encoder-decoder networks are trained and used alone or in combination to deblur the input image. It is an end-to-end approach and introduces no corrugated artifacts as traditional blind deconvolution methods do. We test our approach on different types of pathology specimens and demonstrate great performance on image blur correction and the subsequent improvement on the diagnosis outcome of AI algorithms.

Fourier ptychographic microscopy (FPM) is a computational approach geared towards creating high-resolution and large field-of-view images without mechanical scanning. To acquire color images of histology slides, it often requires sequential acquisitions with red, green, and blue illuminations. The color reconstructions often suffer from coherent artifacts that are not presented in regular incoherent microscopy images. As a result, it remains a challenge to employ FPM for digital pathology applications, where resolution and color accuracy are of critical importance. Here we report a deep learning approach for performing unsupervised image-to-image translation of FPM reconstructions. A cycle-consistent adversarial network with multiscale structure similarity loss is trained to perform virtual brightfield and fluorescence staining of the recovered FPM images. In the training stage, we feed the network with two sets of unpaired images: 1) monochromatic FPM recovery, and 2) color or fluorescence images captured using a regular microscope. In the inference stage, the network takes the FPM input and outputs a virtually stained image with reduced coherent artifacts and improved image quality. We test the approach on various samples with different staining protocols. High-quality color and fluorescence reconstructions validate its effectiveness.

Whole slide imaging (WSI) has moved digital pathology closer to diagnostic practice in recent years. Due to the inherent tissue topography variability, accurate autofocusing remains a critical challenge for WSI and automated microscopy systems. The traditional focus map surveying method is limited in its ability to acquire a high degree of focus points while still maintaining high throughput. Real-time approaches decouple image acquisition from focusing, thus allowing for rapid scanning while maintaining continuous accurate focus. This work reviews the traditional focus map approach and discusses the choice of focus measure for focal plane determination. It also discusses various real-time autofocusing approaches including reflective-based triangulation, confocal pinhole detection, low-coherence interferometry, tilted sensor approach, independent dual sensor scanning, beam splitter array, phase detection, dual-LED illumination, and deep-learning approaches. The technical concepts, merits, and limitations of these methods are explained and compared to those of a traditional WSI system. This review may provide new insights for the development of high-throughput automated microscopy imaging systems that can be made broadly available and utilizable without loss of capacity.

We report an angle-tilted, wavelength-multiplexed ptychographic modulation approach for multispectral lensless on-chip microscopy. In this approach, we illuminate the specimen with lights at 5 wavelengths simultaneously. A prism is added at the illumination path for spectral dispersion. Lightwaves at different wavelengths, thus, hit the specimen at slightly different incident angles, breaking the ambiguities in mixed state ptychographic reconstruction. At the detection path, we place a thin diffuser in-between the specimen and the monochromatic image sensor for encoding the spectral information into 2D intensity measurements. By scanning the sample to different x-y positions, we acquire a sequence of monochromatic images for reconstructing the 5 complex object profiles at the 5 wavelengths. An up-sampling procedure is integrated into the recovery process to bypass the resolution limit imposed by the imager pixel size. We demonstrate a half-pitch resolution of 0.55 microns using an image sensor with 1.85-micron pixel size. We also demonstrate quantitative and high-quality multispectral reconstructions of stained tissue sections for digital pathology applications.

Complex-field imaging is indispensable for numerous applications at wavelengths from X-ray to THz, with amplitude describing transmittance (or reflectivity) and phase revealing intrinsic structure of the target object. Coherent diffraction imaging (CDI) employs iterative phase retrieval algorithms to process diffraction measurements and is the predominant non-interferometric method to image complex fields. However, the working spectrum of CDI is quite narrow, because the diffraction measurements on which it relies require dense array detection with ultra-high dynamic range. Here we report a single-pixel CDI technique that works for a wide waveband. A single-pixel detector instead of an array sensor is employed in the far field for detection. It repeatedly records the DC-only component of the diffracted wavefront scattered from an object as it is illuminated by a sequence of binary modulation patterns. This decreases the measurements' dynamic range by several orders of magnitude. We employ an efficient single-pixel phase-retrieval algorithm to jointly recover the object's 2D amplitude and phase maps from the 1D intensity-only measurements. No a priori object information is needed in the recovery process. We validate the technique's quantitative phase imaging nature using both calibrated phase objects and biological samples, and demonstrate its wide working spectrum with both 488-nm visible light and 980-nm near-infrared light. Our approach paves the way for complex-field imaging in a wider waveband where 2D detector arrays are not available, with broad applications in life and material sciences.

Recent advancements in whole slide imaging (WSI) have moved pathology closer to digital practice. Existing systems require precise mechanical control and the cost is prohibitive for most individual pathologists. Here we report a low-cost and high-throughput WSI system termed OpenWSI. The reported system is built using off-the-shelf components including a programmable LED array, a photographic lens, and a low-cost computer numerical control (CNC) router. Different from conventional WSI platforms, our system performs real-time single-frame autofocusing using color-multiplexed illumination. For axial positioning control, we perform coarse adjustment using the CNC router and precise adjustment using the ultrasonic motor ring in the photographic lens. By using a 20X objective lens, we show that the OpenWSI system has a resolution of ~0.7 microns. It can acquire whole slide images of a 225-mm^2 region in ~2 mins, with throughput comparable to existing high-end platforms. The reported system offers a turnkey solution to transform the high-end WSI platforms into one that can be made broadly available and utilizable without loss of capacity.

Objects classification generally relies on image acquisition and analysis. Real-time classification of high-speed moving objects is challenging, as both high temporal resolution in image acquisition and low computational complexity in objects classification algorithms are required. Here we propose and experimentally demonstrate an approach for real-time moving objects classification without image acquisition. As objects classification algorithms rely on the feature information of objects, we propose to use spatial light modulation to acquire the feature information directly rather than performing image acquisition followed by features extraction. A convolutional neural network is designed and trained to learn the spatial features of the target objects. The trained network can generate structured patterns for spatial light modulation. Using the resulting structured patterns for spatial light modulation, the feature information of target objects can be compressively encoded into a short light intensity sequence. The resulting one-dimensional signal is collected by a single-pixel detector and fed to the convolutional neural network for objects classification. As experimentally demonstrated, the proposed approach can achieve accurate and real-time classification of fast moving objects. The proposed method has potential applications in the fields where fast moving objects classification in real time and for long duration is required.

We report a novel lensless on-chip microscopy platform based on near-field blind ptychographic modulation. In this platform, we place a thin diffuser in between the object and the image sensor for light wave modulation. By blindly scanning the unknown diffuser to different x-y positions, we acquire a sequence of modulated intensity images for quantitative object recovery. Different from previous ptychographic implementations, we employ a unit magnification configuration with a Fresnel number of ~50,000, which is orders of magnitude higher than previous ptychographic setups. The unit magnification configuration allows us to have the entire sensor area, 6.4 mm by 4.6 mm, as the imaging field of view. The ultra-high Fresnel number enables us to directly recover the positional shift of the diffuser in the phase retrieval process, addressing the positioning accuracy issue plagued in regular ptychographic experiments. In our implementation, we use a low-cost, DIY scanning stage to perform blind diffuser modulation. Precise mechanical scanning that is critical in conventional ptychography experiments is no longer needed in our setup. We further employ an up-sampling phase retrieval scheme to bypass the resolution limit set by the imager pixel size and demonstrate a half-pitch resolution of 0.78 micron. We validate the imaging performance via in vitro cell cultures, transparent and stained tissue sections, and a thick biological sample. We show that the recovered quantitative phase map can be used to perform effective cell segmentation of the dense yeast culture. We also demonstrate 3D digital refocusing of the thick biological sample based on the recovered wavefront. The reported platform provides a cost-effective and turnkey solution for large field-of-view, high-resolution, and quantitative on-chip microscopy.

Imaging of biological cells and tissues often relies on fluorescent labels, which offer high contrast with molecular specificity. The use of exogenous labeling agents, however, may alter the normal physiology of the bio-specimens. Complementary to the established fluorescence microscopy, label-free quantitative phase imaging provides an objective morphological measurement tool for bio-specimens and is free of variability introduced by contrast agents. Here we report a simple and low-cost microscope add-on, termed Ptychographic Modulation Engine (PME), for super-resolution quantitative phase imaging. In this microscope add-on module, we attach a diffuser to a 3D-printed holder that can be mechanically moved to different x-y positions. We then use two vibrational motors to introduce random positional shifts to the diffuser. The add-on module can be placed between the objective lens and the specimen in most existing microscope platforms. Thanks to the diffuser modulation process, the otherwise inaccessible high-resolution object information can now be encoded into the captured images. In the ptychographic phase retrieval process, we jointly recover the complex object wavefront, the complex diffuser profile, and the unknown positional shifts of the diffuser. We demonstrate a 4-fold resolution gain over the diffraction limit of the employed 2X objective lens. We also test our approach for in-vivo cell imaging, where we are able to adjust the focus after the data has been captured. The reported microscope add-on provides a turnkey solution for super-resolution quantitative phase imaging. It may find applications in label-free bio-imaging where both large field-of-view and high resolution are needed.

Background: The use of whole slide imaging (WSI) for digital pathology has recently been cleared for primary diagnosis in the US. A conventional WSI system scans the tissue slide to different positions and acquires the digital images. In a typical implementation, a focus map is created prior to the scanning process, leading to significant overhead time and a necessity for high positional accuracy of the mechanical system. The resulting cost of WSI system is often prohibitive for frozen section procedure during surgery. Methods: We report a novel WSI scheme based on a programmable LED array for sample illumination. In between two regular brightfield image acquisitions, we acquire one additional image by turning on a red and a green LED for color multiplexed illumination. We then identify the translational shift of the red- and green-channel images by maximizing the image mutual information or cross-correlation. The resulting translational shift is used for dynamic focus correction in the scanning process. Since we track the differential focus during adjacent acquisitions, there is no positional repeatability requirement in our scheme. Results: We demonstrate a prototype WSI platform with a mean focusing error of ~0.3 microns. Different from previous implementations, this prototype platform requires no focus map surveying, no secondary camera or additional optics, and allows for continuous sample motion in the focus tracking process. Conclusions: A programmable LED array can be used for color-multiplexed single-shot autofocusing in WSI. The reported scheme may enable the development of cost-effective WSI platforms without positional repeatability requirement. It may also provide a turnkey solution for other high-content microscopy applications.

Digital aberration measurement and removal play a prominent role in computational imaging platforms aimed at achieving simple and compact optical arrangements. A recent important class of such platforms is Fourier ptychography, which is geared towards efficiently creating gigapixel images with high resolution and large field of view (FOV). In current FP implementations, pupil aberration is often recovered at each small segment of the entire FOV. This reconstruction strategy fails to consider the field-dependent nature of the optical pupil. Given the power series expansion of the wavefront aberration, the spatially varying pupil can be fully characterized by tens of coefficients over the entire FOV. With this observation, we report a Full-field Fourier Ptychography (FFP) scheme for rapid and robust aberration metrology. The meaning of 'full-field' in FFP is referred to the recovering of the 'full-field' coefficients that govern the field-dependent pupil over the entire FOV. The optimization degrees of freedom are at least two orders of magnitude lower than the previous implementations. We show that the image acquisition process of FFP can be completed in ~1s and the spatially varying aberration of the entire FOV can be recovered in ~35s using a CPU. The reported approach may facilitate the further development of Fourier ptychography. Since no moving part or calibration target is needed in this approach, it may find important applications in aberration metrology. The derivation of the full-field coefficients and its extension for Zernike modes also provide a general tool for analyzing spatially varying aberrations in computational imaging systems.

We report a new coherent imaging technique, termed ptychographic structured modulation (PSM), for quantitative super-resolution microscopy. In this technique, we place a thin diffuser (i.e., a scattering lens) in between the sample and the objective lens to modulate the complex light waves from the object. The otherwise inaccessible high-resolution object information can thus be encoded into the captured images. We then employ a ptychographic phase retrieval process to jointly recover the exit wavefront of the complex object and the unknown diffuser profile. Unlike the illumination-based super-resolution approach, the recovered image of our approach depends upon how the complex wavefront exits the sample - not enters it. Therefore, the sample thickness becomes irrelevant during reconstruction. After recovery, we can propagate the super-resolution complex wavefront to any position along the optical axis. We validate our approach using a resolution target, a quantitative phase target, a two-layer sample, and a thick PDMS sample. We demonstrate a 4.5-fold resolution gain over the diffraction limit. We also show that a 4-fold resolution gain can be achieved with as few as ~30 images. The reported approach may provide a quantitative super-resolution strategy for coherent light, X-ray, and electron imaging.

We report a compact, cost-effective and field-portable lensless imaging platform for quantitative microscopy. In this platform, the object is placed on top of an image sensor chip without using any lens. We use a low-cost galvo scanner to rapidly scan an unknown laser speckle pattern on the object. To address the positioning repeatability and accuracy issues, we directly recover the positional shifts of the speckle pattern based on the phase correlation of the captured images. To bypass the resolution limit set by the imager pixel size, we employ a sub-sampled ptychographic phase retrieval process to recover the complex object. We validate our approach using a resolution target, a phase target, and a biological sample. Our results show that accurate, high-quality complex images can be obtained from a lensless dataset with as few as ~10 images. We also demonstrate the reported approach to achieve a 6.4 mm by 4.6 mm field of view and a half pitch resolution of 1 miron. The reported approach may provide a quantitative lensless imaging strategy for addressing point-of-care, global-health, and telemedicine related challenges.

Achieving high spatial resolution is the goal of many imaging systems. Designing a high-resolution lens with diffraction-limited performance over a large field of view remains a difficult task in imaging system design. On the other hand, creating a complex speckle pattern with wavelength-limited spatial features is effortless and can be implemented via a simple random diffuser. With this observation and inspired by the concept of near-field ptychography, we report a new imaging modality, termed near-field Fourier ptychography, for tackling high-resolution imaging challenges in both microscopic and macroscopic imaging settings. The meaning of 'near-field' is referred to placing the object at a short defocus distance with a large Fresnel number. In our implementations, we project a speckle pattern with fine spatial features on the object instead of directly resolving the spatial features via a high-resolution lens. We then translate the object (or speckle) to different positions and acquire the corresponding images using a low-resolution lens. A ptychographic phase retrieval process is used to recover the complex object, the unknown speckle pattern, and the coherent transfer function at the same time. In a microscopic imaging setup, we use a 0.12 numerical aperture (NA) lens to achieve a NA of 0.85 in the reconstruction process. In a macroscale photographic imaging setup, we achieve ~7-fold resolution gain using a photographic lens. The final achievable resolution is not determined by the collection optics. Instead, it is determined by the feature size of the speckle pattern. The reported imaging modality can be employed in light, coherent X-ray, and transmission electron imaging systems to increase resolution and provide quantitative absorption and phase contrast of the object., Comment: 15 pages, 14 figures

Structured illumination has been widely used for optical sectioning and 3D surface recovery. In a typical implementation, multiple images under non-uniform pattern illumination are used to recover a single object section. Axial scanning of the sample or the objective lens is needed for acquiring the 3D volumetric data. Here we demonstrate the use of axially-shifted pattern illumination (asPI) for virtual volumetric confocal imaging without axial scanning. In the reported approach, we project illumination patterns at a tilted angle with respect to the detection optics. As such, the illumination patterns shift laterally at different z sections and the sample information at different z-sections can be recovered based on the captured 2D images. We demonstrate the reported approach for virtual confocal imaging through a diffusing layer and underwater 3D imaging through diluted milk. We show that we can acquire the entire confocal volume in ~1s with a throughput of 420 megapixels per second. Our approach may provide new insights for developing confocal light ranging and detection systems in degraded visual environments.

Fourier ptychography is a recently developed imaging approach for large field-of-view and high-resolution microscopy. Here we model the Fourier ptychographic forward imaging process using a convolution neural network (CNN) and recover the complex object information in the network training process. In this approach, the input of the network is the point spread function in the spatial domain or the coherent transfer function in the Fourier domain. The object is treated as 2D learnable weights of a convolution or a multiplication layer. The output of the network is modeled as the loss function we aim to minimize. The batch size of the network corresponds to the number of captured low-resolution images in one forward / backward pass. We use a popular open-source machine learning library, TensorFlow, for setting up the network and conducting the optimization process. We analyze the performance of different learning rates, different solvers, and different batch sizes. It is shown that a large batch size with the Adam optimizer achieves the best performance in general. To accelerate the phase retrieval process, we also discuss a strategy to implement Fourier-magnitude projection using a multiplication neural network model. Since convolution and multiplication are the two most-common operations in imaging modeling, the reported approach may provide a new perspective to examine many coherent and incoherent systems. As a demonstration, we discuss the extensions of the reported networks for modeling single-pixel imaging and structured illumination microscopy (SIM). 4-frame resolution doubling is demonstrated using a neural network for SIM. We have made our implementation code open-source for the broad research community.

Whole slide imaging (WSI) has recently been cleared for primary diagnosis in the US. A critical challenge of WSI is to perform accurate focusing in high speed. Traditional systems create a focus map prior to scanning. For each focus point on the map, sample needs to be static in the x-y plane and axial scanning is needed to maximize the contrast. Here we report a novel focus map surveying method for WSI. The reported method requires no axial scanning, no additional camera and lens, works for stained and transparent samples, and allows continuous sample motion in the surveying process. It can be used for both brightfield and fluorescence WSI. By using a 20X, 0.75 NA objective lens, we demonstrate a mean focusing error of ~0.08 microns in the static mode and ~0.17 microns in the continuous motion mode. The reported method may provide a turnkey solution for most existing WSI systems for its simplicity, robustness, accuracy, and high-speed. It may also standardize the imaging performance of WSI systems for digital pathology and find other applications in high-content microscopy such as DNA sequencing and time-lapse live-cell imaging.

Single-pixel imaging is an innovative imaging scheme and has received increasing attentions in recent years. It is applicable to imaging at non-visible wavelengths and imaging under low light conditions. However, single-pixel imaging has once encountered problems of low reconstruction quality and long data-acquisition time. This situation has been changed thanks to the developments of Hadamard single-pixel imaging (HSI) and Fourier single-pixel imaging (FSI). Both techniques are able to achieve high-quality and efficient imaging, remarkably improving the applicability of single-pixel imaging scheme. In this paper, we compare the performances of HSI and FSI with theoretical analysis and experiments. The results show that FSI is more efficient than HSI while HSI is more noise-robust than FSI. Our work may provide a guideline for researchers to choose suitable single-pixel imaging technique for their applications.

We report the development of a multichannel microscopy for whole-slide multiplane, multispectral, and phase imaging. We use trinocular heads to split the beam path into 6 independent channels and employ a camera array for parallel data acquisition, achieving a maximum data throughput of ~1 gigapixel per second. To perform single-frame rapid autofocusing, we place two near-infrared LEDs at the back focal plane of the condenser lens to illuminate the sample from two different incident angles. A hot mirror is used to direct the near-infrared light to an autofocusing camera. For multiplane whole slide imaging (WSI), we acquire 6 different focal planes of a thick specimen simultaneously. For multispectral WSI, we relay the 6 independent image planes to the same focal position and simultaneously acquire information at 6 spectral bands. For whole-slide phase imaging, we acquire images at 3 focal positions simultaneously and use the transport-of-intensity equation to recover the phase information. We also provide an open-source design to further increase the number of channels from 6 to 15. The reported platform provides a simple solution for multiplexed fluorescence imaging and multimodal WSI. Acquiring an instant focal stack without z-scanning may also enable fast 3D dynamic tracking of various biological samples., Comment: 6 pages, 8 figures

Fourier ptychography (FP) is a recently proposed computational imaging technique for high space-bandwidth product imaging. In real setups such as endoscope and transmission electron microscope, the common sample motion largely degrades the FP reconstruction and limits its practicability. In this paper, we propose a novel FP reconstruction method to efficiently correct for unknown sample motion. Specifically, we adaptively update the sample's Fourier spectrum from low spatial-frequency regions towards high spatial-frequency ones, with an additional motion recovery and phase-offset compensation procedure for each sub-spectrum. Benefiting from the phase retrieval redundancy theory, the required large overlap between adjacent sub-spectra offers an accurate guide for successful motion recovery. Experimental results on both simulated data and real captured data show that the proposed method can correct for unknown sample motion with its standard deviation being up to 10% of the field-of-view scale. We have released our source code for non-commercial use, and it may find wide applications in related FP platforms such as endoscopy and transmission electron microscopy.

Recently Fourier Ptychography (FP) has attracted great attention, due to its marked effectiveness in leveraging snapshot numbers for spatial resolution in large field-of-view imaging. To acquire high signal-to-noise-ratio (SNR) images under angularly varying illuminations for subsequent reconstruction, FP requires long exposure time, which largely limits its practical applications. In this paper, based on the recently reported Wirtinger flow algorithm, we propose an iterative optimization framework incorporating phase retrieval and noise relaxation together, to realize FP reconstruction using low SNR images captured under short exposure time. Experiments on both synthetic and real captured data validate the effectiveness of the proposed reconstruction method. Specifically, the proposed technique could save around 80% exposure time to achieve similar retrieval accuracy compared to the conventional FP. Besides, we have released our source code for non-commercial use.