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Pengming Song

Publications and source records attributed to Pengming Song.

At least 19 recordsLinked to original sources

Deep-ultraviolet ptychographic pocket-scope (DART): mesoscale lensless molecular imaging with label-free spectroscopic contrast

The mesoscale characterization of biological specimens has traditionally required compromises between resolution, field-of-view, depth-of-field, and molecular specificity, with most approaches relying on external labels. Here we present the Deep-ultrAviolet ptychogRaphic pockeT-scope (DART), a handheld platform that transforms label-free molecular imaging through intrinsic deep-ultraviolet spectroscopic contrast. By leveraging biomolecules' natural absorption fingerprints and combining them with lensless ptychographic microscopy, DART resolves down to 308-nm linewidths across centimeter-scale areas while maintaining millimeter-scale depth-of-field. The system's virtual error-bin methodology effectively eliminates artifacts from limited temporal coherence and other optical imperfections, enabling high-fidelity molecular imaging without lenses. Through differential spectroscopic imaging at deep-ultraviolet wavelengths, DART quantitatively maps nucleic acid and protein distributions with femtogram sensitivity, providing an intrinsic basis for explainable virtual staining. We demonstrate DART's capabilities through molecular imaging of tissue sections, cytopathology specimens, blood cells, and neural populations, revealing detailed molecular contrast without external labels. The combination of high-resolution molecular mapping and broad mesoscale imaging in a portable platform opens new possibilities from rapid clinical diagnostics, tissue analysis, to biological characterization in space exploration.

physics.optics

Ptychographic non-line-of-sight imaging for depth-resolved visualization of hidden objects

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.

eess.IV

Ptycho-endoscopy on a lensless ultrathin fiber bundle tip

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.

physics.optics

Spatially-coded Fourier ptychography: flexible and detachable coded thin films for quantitative phase imaging with uniform phase transfer characteristics

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.

physics.optics

Lensless polarimetric coded ptychography for high-resolution, high-throughput gigapixel birefringence imaging on a chip

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.

physics.optics

Blood-coated sensor for high-throughput ptychographic cytometry on a Blu-ray disc

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.

physics.ins-det

Lensless coherent diffraction imaging based on spatial light modulator with unknown modulation curve

Lensless imaging is a popular research field for the advantages of small size, wide field-of-view and low aberration in recent years. However, some traditional lensless imaging methods suffer from slow convergence, mechanical errors and conjugate solution interference, which limit its further application and development. In this work, we proposed a lensless imaging method based on spatial light modulator (SLM) with unknown modulation curve. In our imaging system, we use SLM to modulate the wavefront of object, and introduce the ptychographic scanning algorithm that is able to recover the complex amplitude information even the SLM modulation curve is inaccurate or unknown. In addition, we also design a split-beam interference experiment to calibrate the modulation curve of SLM, and using the calibrated modulation function as the initial value of the expended ptychography iterative engine (ePIE) algorithm can improve the convergence speed. We further analyze the effect of modulation function, algorithm parameters and the characteristics of the coherent light source on the quality of reconstructed image. The simulated and real experiments show that the proposed method is superior to traditional mechanical scanning methods in terms of recovering speed and accuracy, with the recovering resolution up to 14 um.

physics.optics

Ptychographic sensor for large-scale lensless microbial monitoring with high spatiotemporal resolution

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.

physics.ins-det

Resolution-enhanced parallel coded ptychography for high-throughput optical imaging

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.

physics.optics

Optofluidic ptychography on a chip

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.

physics.ins-det

High-throughput lensless whole slide imaging via continuous height-varying modulation of tilted sensor

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.

eess.IV

Bypassing the resolution limit of diffractive zone plate optics via rotational Fourier ptychography

Diffractive zone plate optics uses a thin micro-structure pattern to alter the propagation direction of the incoming light wave. It has found important applications in extreme-wavelength imaging where conventional refractive lenses do not exist. The resolution limit of zone plate optics is determined by the smallest width of the outermost zone. In order to improve the achievable resolution, significant efforts have been devoted to the fabrication of very small zone width with ultrahigh placement accuracy. Here, we report the use of a diffractometer setup for bypassing the resolution limit of zone plate optics. In our prototype, we mounted the sample on two rotation stages and used a low-resolution binary zone plate to relay the sample plane to the detector. We then performed both in-plane and out-of-plane sample rotations and captured the corresponding raw images. The captured images were processed using a Fourier ptychographic procedure for resolution improvement. The final achievable resolution of the reported setup is not determined by the smallest width structures of the employed binary zone plate; instead, it is determined by the maximum angle of the out-of-plane rotation. In our experiment, we demonstrated 8-fold resolution improvement using both a resolution target and a titanium dioxide sample. The reported approach may be able to bypass the fabrication challenge of diffractive elements and open up new avenues for microscopy with extreme wavelengths.

physics.optics

Virtual brightfield and fluorescence staining for Fourier ptychography via unsupervised deep learning

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.

eess.IV

Autofocusing technologies for whole slide imaging and automated microscopy

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.

physics.med-ph

Super-resolved multispectral lensless microscopy via angle-tilted, wavelength-multiplexed ptychographic modulation

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.

physics.optics

OpenWSI: a low-cost, high-throughput whole slide imaging system via single-frame autofocusing and open-source hardware

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.

eess.IV

Wide-field, high-resolution lensless on-chip microscopy via near-field blind ptychographic modulation

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.

eess.IV

Ptychographic modulation engine (PME): a low-cost DIY microscope add-on for coherent super-resolution imaging

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.

physics.ins-det