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Maciej Trusiak

Publications and source records attributed to Maciej Trusiak.

At least 19 recordsLinked to original sources

Hybrid spectral-spatial domain registration for nanometric tracking in digital in-line holographic microscopy

Digital in-line holographic microscopy enables label-free tracking and metrology, but achieving nanometric sub-pixel displacement accuracy over a wide capture range remains challenging. Frequency-domain registration based on the discrete Fourier transform (DFT) is globally stable and tolerant to large shifts, yet it suffers from sub-pixel quantization and interpolation artifacts that limit precision near zero displacement. In contrast, spatial-gradient refinement such as Lucas-Kanade (LK) can reach very high sub-pixel accuracy, but it is strongly initialization-limited and prone to divergence outside a narrow convergence basin. Here we propose a Hybrid Spectral-Spatial Domain (HSSD) framework that resolves this trade-off by combining the global robustness of a DFT-based coarse estimator with LK refinement of the residual motion. The DFT stage provides reliable initialization and substantially extends the capture range, while the LK stage suppresses the precision floor by reducing interpolation and quantization errors characteristic of standalone frequency-domain methods. We validate HSSD using numerical simulations and experiments in a transmission in-line holographic imaging system, achieving nanoprecision displacement measurement and stable tracking across a wide range of displacements and defocusing conditions. This hybrid strategy enables reliable nanometric localization in regimes where standalone DFT or LK methods either fail to converge or saturate in accuracy.

physics.optics

Lateral shearing optical diffraction tomography of brain organoid with reduced spatial coherence

Optical diffraction tomography (ODT) is a powerful technique for quantitative, label-free reconstruction of the three-dimensional refractive index (RI) distribution of biological samples. While ODT is well established for imaging thin, weakly scattering samples, it encounters significant challenges when applied to heterogeneous, strongly scattering thick samples such as tissues and organoids. In this work, a novel common-path interferometric approach to ODT is presented, specifically designed for the RI reconstruction of heterogeneous and highly scattering samples at high temporal stability. The proposed technique, termed lateral shearing (LS)-ODT, incorporates partial lateral shearing off-axis interferometry to suppress the effects of multiple scattering, similar to the mechanism in differential interference contrast (DIC) microscopy, which is widely used for imaging thick specimens. Additionally, the LS-ODT system uses dynamic speckle illumination to enhance both spatial phase and RI sensitivity compared to laser-based ODT systems. The effectiveness of this method is demonstrated through experiments on a cell phantom. Its robustness and accuracy are further validated across a wide range of samples, including mouse kidney tissue sections and brain organoids derived from human induced pluripotent stem cells (iPSCs), in both thin and thick sections. Furthermore, correlative fluorescence and RI tomography of the organoids highlights the potential of LS-ODT to enhance and support a broad spectrum of biomedical studies, particularly in the field of histology.

physics.optics

High-Sensitivity, High-Throughput Double Sagnac Lateral Shearing Quantitative Phase Microscopy and Tomography with Pseudo-Thermal Illumination

Quantitative phase microscopy (QPM) enables label-free measurement of local optical path length variations, providing critical insight into the structure and dynamics of transparent biological specimens. Here, a highly sensitive lateral shearing QPM (LS-QPM) system is presented, based on a novel double Sagnac common-path interferometric configuration combined with pseudo-thermal illumination. The pseudo-thermal light source plays a central role in enhancing spatial phase sensitivity by suppressing coherent noise and speckle artifacts, while maintaining sufficient temporal coherence to generate high-density interference fringes, thereby enabling robust single-shot phase retrieval. In addition, the double Sagnac architecture introduces an inherently stable common-path geometry, significantly enhancing temporal phase stability. Unlike conventional quadriwave lateral shearing interferometry (QWLSI)-based QPM systems, which typically suffer from a trade-off between spatial resolution and field of view (FOV), the proposed approach enables simultaneous achievement of diffraction-limited resolution and a large FOV. Experimental validation using calibrated polystyrene beads demonstrates accurate and spatially uniform phase reconstruction across the entire imaging area. Further, the system's capability for biological imaging is demonstrated through experiments on fixed and live HeLa cells, where subcellular features and dynamic processes are captured in a label-free manner. Furthermore, volumetric imaging of embedded bead samples highlights the potential of the approach for three-dimensional phase tomography. The lateral shearing mechanism, analogous to differential interference contrast (DIC), improves robustness to multiple scattering, indicating strong potential for future applications in imaging thick and heterogeneous samples.

physics.optics

YOSO: single-frame Gerchberg-Saxton phase retrieval with AI-based data augmentation for in-line holography

We present YOSO (You Only Shot Once), a single-frame phase retrieval framework for digital in-line holographic microscopy (DIHM) in which supervised deep learning is used to numerically generate an additional hologram corresponding to different defocus distance, creating a so-called multi-height dataset, which is then conventionally processed with a well-established Gerchberg-Saxton (GS) algorithm. YOSO is trained on computer-generated data derived from natural images, enabling strong generalization. The selected multi-scale ResNet architecture enables rapid training in under two hours on a mid-range workstation, which is done only once, enabling efficient inference thereafter. We further show that YOSO network can process inputs of varying spatial dimensions, allowing training on small inputs and direct inference on full-sized holograms while bypassing patch-and-stitch procedure. A further advantage of YOSO is its physics-consistent hologram padding, which replaces conventional zero or edge-value padding with a physically grounded approach compatible with the GS framework. The YOSO framework is tested on various systems (lens-based and lensless DIHM) and diverse samples: a resolution test target, adherent and suspended biological cells, and a mouse brain slice. The results show that YOSO is compatible with 3D objects and correctly recovers defocused object wave features, enabling holographic postprocessing such as numerical refocusing. The results of this work are available publicly as software for end-to-end implementation.

physics.optics

Near-infrared lensless holographic microscopy on a visible sensor enables label-free high-throughput imaging in strong scattering

Lensless digital holographic microscopy (LDHM) relies on interference between an unscattered reference wave and a weakly scattered object wave - an assumption that rapidly fails in turbid samples under multiple scattering. To overcome this limitation, we present near-infrared LDHM (NIR-LDHM), a in-line holographic platform that operates up to the silicon cutoff (~1100 nm) using a conventional board-level CMOS sensor designed for visible (VIS) imaging. Using tissue-mimicking milk scattering layers and calibrated resolution targets, we quantify reconstruction performance versus wavelength, scattering strength, and sample-sensor distance. NIR-LDHM maintains resolvable features through scattering layers up to ~1.4 mm, whereas visible regime fails to resolve features below ~350 um. Importantly, despite a detector quantum efficiency of only 0.19% at 1100 nm, robust reconstructions are obtained under low-photon-budget conditions. We further identify advantageous mechanism by which increasing the sample-sensor distance from ~3 to 12 mm improves lateral resolution by twofold under strong scattering. Finally, we demonstrate wide-field, label-free amplitude-phase imaging of uncleared mouse tissues, resolving internal structure in brain and liver slices up to ~250 um and ~60 um, respectively. By extending lensless complex-field microscopy into strongly scattering regimes with minimal hardware changes, this work has relevance to computational imaging through complex media and biophotonics.

physics.optics

The mixture of glycerin with tartrazine: a solution to reversibly increase tissue transparency for in vitro quantitative phase imaging

Thick tissue sections strongly scatter and absorb light, which limits transmission-based label-free examination via quantitative phase imaging (QPI) modalities. Here we introduce a simple, room-temperature optical clearing medium - glycerol and tartrazine solution (GTS; 60 percent glycerol, 10 percent tartrazine) - that increases the transparency of 50-80 micron murine liver and kidney slices while preserving tissue morphology and enabling rapid, label-free quantitative phase imaging. Using Fourier ptychographic microscopy (FPM) and lensless digital holographic microscopy with pixel super-resolution (LDHM-PSR), we demonstrate markedly improved high-throughput visualization of microstructural features after GTS immersion compared with phosphate-buffered saline (PBS). The improvement is confirmed quantitatively by a significant increase in the one-pixel-lag autocorrelation of phase-gradient values, indicating reduced scattering-driven phase artifacts and enhanced structural continuity in reconstructed phase maps. We further show that GTS is stable for months at room temperature, provides instant yet durable clearing that improves over time, and can be removed by brief PBS washing, enabling downstream multimodal analyses on the same specimen. Finally, we benchmark GTS against the state-of-the-art clearing agent Ce3D, highlighting GTS as a low-cost, safe, operationally straightforward alternative with strong potential for routine, high-throughput QPI-based histopathology workflows.

physics.optics

DeepBessel: deep learning-based full-field vibration profilometry using single-shot time-averaged interference microscopy

Full-field vibration profilometry is essential for dynamic characterizing microelectromechanical systems (MEMS/MOEMS). Time-averaged interferometry (TAI) encodes spatial information about MEMS or MOEMS vibration amplitude in the interferogram's amplitude modulation using Bessel function (besselogram). Classical approaches for interferogram analysis are specialized for cosine function fringe patterns and therefore introduce reconstruction errors for besselogram decoding. This paper presents the DeepBessel: a deep learning-based approach for single-shot TAI interferogram analysis. A convolutional neural network (CNN) was trained using synthetic data, where the input consisted of besselograms, and the output corresponded to the underlying vibration amplitude distribution. Numerical validation and experimental testing demonstrated that DeepBessel significantly reduces reconstruction errors compared to the state-of-the-art approaches, e.g., Hilbert Spiral Transform (HST) method. The proposed network effectively mitigates errors caused by the mismatch between the Bessel and cosine functions. The results indicate that deep learning can improve the accuracy of full-field vibration measurements, offering new possibilities for optical metrology in MEMS or MOEMS applications.

physics.optics

High Space-bandwidth Product Label-free Examination of iPSC-derived Brain Organoids via Fourier Ptychographic Microscopy

Fourier ptychographic microscopy (FPM) is a promising quantitative phase imaging technique that enables high-resolution, label-free imaging over a large field-of-view. Here, we present the first application of FPM for the quantitative analysis of human brain organoid slices, providing a powerful, cost-effective, and label-free enhancement to the current gold-standard fluorescence microscopy. Brain organoids, prepared as thin (5 micrometer) slices, were imaged with a custom-built FPM system consisting of a standard light microscope (4x, 0.2 NA objective) and a 7x7 LED array. This configuration achieved a synthetic numerical aperture of 0.54 and a spatial resolution of approximately 488 nm across an area of 2.077 x 3.65 mm. Fluorescence microscopy was used in parallel for neurons, astrocytes, and nuclei labeling, providing rich fluorescence imaging. Moreover, we designed an automated method to merge classical resolution fluorescence images to visualize the whole brain organoid and align it with the numerically increased space-bandwidth product FPM image. The provided alignment method enables rich phase-fluorescence correlative imaging. Based on the segmentation performed on the stitched fluorescence images, we devised a quantitative phase analysis revealing a higher mean optical thickness of the nuclei versus astrocytes and neurons. Notably, nuclei located in neurogenic regions consistently exhibited significantly higher phase values (optical path difference) compared to nuclei elsewhere, suggesting cell-type-specific biophysical signatures. The label-free, quantitative, and high-throughput capabilities of the FPM approach demonstrated here make it a powerful and accessible tool for future structural and functional studies of whole-section brain organoid development and disease modeling studies.

physics.med-ph

Low-dose Chemically Specific Bioimaging via Deep-UV Lensless Holographic Microscopy on a Standard Camera

Deep-ultraviolet (DUV) microscopy can provide label-free biochemical contrast by exploiting the intrinsic absorption of nucleic acids, proteins and lipids, offering chemically specific morphological information that complements structural optical thickness contrast from phase-sensitive imaging. However, existing DUV microscopes typically rely on specialized optics and DUV-sensitive cameras, which restrict field of view, increase system complexity and cost, and often require high illumination doses that risk photodamage. Here, we report a low-dose deep-UV lensless holographic microscopy platform that uses standard board-level CMOS sensors designed for visible light, eliminating all imaging optics and dedicated DUV detectors. Our system achieves large field-of-view (up to 116 mm2) DUV imaging with low illumination and label-free phase and chemically specific amplitude contrast. A specialized defocus/wavelength diverse pixel super-resolution reconstruction with total-variation regularization and robust autofocusing halves the effective sensor pixel pitch and yields down to 870 nm lateral resolution. We demonstrate chemically specific, label-free bioimaging on challenging specimens, including Saccharomyces cerevisiae, extracellular vesicles and unstained mouse liver tissue. In liver sections, imaging at λ = 330 nm reveals lipid- and retinoid-rich accumulations that co-localize with Oil Red O staining, enabling label-free identification of hepatic stellate (Ito) cells. This combination of low-dose operation, chemically specific contrast and standard CMOS hardware establishes DUV lensless holographic microscopy as a practical and scalable route to high-content submicron-resolution whole-slide preparation-free bioimaging without exogenous labels.

physics.optics

Bayesian inference for precise and uncertainty-quantified single-shot widefield interferometric geometrical nanometrology

Advanced geometrical nanometrology is critical for process control in semiconductor manufacturing, supporting applications in, e.g., photonic integrated circuits, nanoelectronics, and emerging quantum and optoelectronic technologies. Widefield interferometric approach provide a cost-effective, non-destructive solution for characterizing semiconductor optical waveguides, which are fundamental to nanophotonic devices. This work presents a Bayesian inference framework, implemented using Dynamic Nested Sampling, for estimating geometric parameters - such as width and height - of a semiconductor optical waveguide from a single widefield interferogram. The proposed framework reduces the need of leveraging near field scanning microscopy methods for measurements. The notable advantage is that Bayesian statistics not only provide the estimated parameter values but also quantify the uncertainty of the inference results and the fitness of the used model. The proposed full-field, single-shot interferometric approach, supported by Bayesian-based data analysis, achieves high accuracy and sensitivity - down to successful measurement of 8 nm rib waveguide - while remaining resilient to noise. Thus, the demonstrated methodology provides a cost-effective, robust, and scalable tool for semiconductor fabrication monitoring and process verification, as confirmed by both numerical simulations and experimental validation on optical waveguides. This method contributes to high-precision nanometrology by integrating advanced statistical modeling and inference techniques.

physics.optics

Doubling the field of view and eliminating the replica overlap problem in common-path shearing quantitative phase imaging

Quantitative phase imaging (QPI) enables label-free, high-contrast visualization of transparent specimens, but its common implementation in off-axis digital holographic microscopy (DHM) requires a separate reference beam, which increases system complexity and sensitivity to noise and vibrations. Common-path shearing DHMs overcome these drawbacks by eliminating the reference arm, yet they suffer from sheared object beam (replica) overlap, as both interfering sheared beams traverse the sample and generate superimposed phase images. This limits their use to sparse objects only. Here we introduce R2D-QPI, a method that numerically decouples object and replica fields of view through controlled shear scanning. The method analytically separates overlapped phase images and effectively doubles the imaged area, requiring only two measurements. We experimentally validate the approach on a phase resolution test target, yeast cells, and human thyroid tissue slices, demonstrating accurate reconstruction even in highly confluent samples with strong object-replica overlap. The results establish R2D-QPI as a robust and versatile solution for common-path QPI, enabling wide-field, label-free phase imaging with minimal data acquisition and strong potential for applications in biological and medical microscopy.

physics.optics

OSI-flex: Optimization-Based Shearing Interferometry for Joint Phase and Shear Estimation Using a Flexible Open-Source Framework

Shearing interferometry is a common-path quantitative phase imaging technique in which an object beam interferes with a laterally shifted replica of itself, providing high temporal stability, reduced sensitivity to environmental noise, compact design, and compatibility with partially coherent illumination that suppresses coherence-related artifacts. Its principal limitation, however, is that it yields only sheared phase-difference measurements rather than the absolute phase, thereby requiring additional reconstruction step. In this work, we introduce OSI-flex, a flexible, open-source computational framework for quantitative phase reconstruction from sheared phase-difference measurements. The method leverages modern machine learning tools, namely automatic differentiation and the advanced ADAM (Adaptive Moment Estimation) optimizer. The method simultaneously estimates the phase and shear values, enabling it to adapt to experimental conditions where the shear cannot be precisely determined. Because defining shear value is inherently difficult in most systems, yet crucial for effective phase reconstruction, this joint optimization leads to robust and reliable phase retrieval. OSI-flex is highly versatile, supporting arbitrary numbers, magnitudes, and orientations of shear vectors. While optimal reconstruction is achieved with two orthogonal shears, the inclusion of regularization - specifically total variation minimization and sign constraint - enables OSI-flex to remain effective with nonorthogonal or even single-shear measurements. Moreover, OSI-flex accommodates a wide range of shear magnitudes, from subpixel (differential configuration) to several dozen pixels (semi-total shear configuration). Validation with simulations and experimental data confirms quantitative accuracy on calibrated phase objects and demonstrates robustness with 3D-printed cell phantom and follicular thyroid cells.

physics.optics

Speckle suppression in digital in-line holographic microscopy through liquid crystal dynamic scattering

We demonstrate speckle noise reduction in an in-line holographic imaging system using a Zwitterion-doped liquid crystal dynamic scatterer (LCDS) cell diffuser. Integrated into a minimally modified bright-field microscope, the LCDS actively modulates system's spatial coherence. The proposed solution suppresses coherent artifacts without introducing bulky moving parts, while enhancing image resolution and preserving overall system simplicity. Quantitative performance tested on a phase and amplitude test targets, as well as phase-amplitude biological sample, shows significant noise reduction and methods versatility. Though validated in a holographic in-line setup, the approach is applicable to other imaging techniques requiring compact, vibration-free speckle suppression.

physics.optics

Gigavoxel-Scale Multiple-Scattering-Aware Lensless Holotomography

Holotomography (HT) has revolutionized quantitative label-free 3D imaging, yet conventional lens-based implementations are fundamentally constrained in field-of-view (FOV) and imaging depth, limiting their utility for critical high-throughput applications in material and life sciences. Lensless HT (LHT) offers a promising alternative for large-volume examination, however existing approaches fail to accurately reconstruct highly scattering samples over extended depths, which remains a critical challenge in optical imaging field. Here, we introduce a gigavoxel-scale, multiple-scattering-aware LHT with a large FOV (surpassing 0.6 cm2), millimeter-scale axial range and pixel level (~2.4 micron) resolution. Our approach leverages a multi-wavelength, oblique-illumination hologram reconstruction and a robust, automatic illumination angle calibration, which are necessary for precise large-volume 3D holographic reconstruction. Moreover, we propose optimization-driven multi-slice tomographic framework to accurately capture multiple-scattering effects outperforming first order Born/Rytov-based inversions. To rigorously validate our method, we reconstruct bespoke multi-layer two-photon polymerized test structure over a 1.7 mm imaging depth and 25 mm2 FOV, yielding an unprecedented 3D space-bandwidth product exceeding a gigavoxel level. Furthermore, we demonstrate for the first time on-chip label-free imaging of entire 500-micron-thick tissue slice of optically-cleared mouse brain. With the proposed method, we aim to unlock powerful new capabilities for large-scale, quantitative, label-free 3D imaging across biomedicine, neuroscience, material sciences and beyond.

physics.optics

Fourier ptychographic microscopy aided with transport of intensity equation for robust full phase spectrum reconstruction

Fourier ptychographic microscopy (FPM) is a pivotal computational imaging technique that achieves phase and amplitude reconstruction with high resolution and wide field of view, using low numerical aperture objectives and LED array illumination. Despite its unique strengths, FPM remains fundamentally limited in retrieving low spatial frequency phase information due to the absence of phase encoding in all brightfield illumination angles. To overcome this, we present a novel hybrid approach that combines FPM with the transport of intensity equation (TIE), enabling accurate, full-spectrum phase retrieval without compromising system simplicity. Our method extends standard FPM acquisitions with a single additional on-axis defocused image, from which low-frequency phase components are reconstructed via TIE method, employing large defocus distance to suppress low-frequency artifacts and enhance robustness to intensity noise. To additionally compensate for defocus-induced magnification variations caused by spherical wavefront illumination, we employ an affine transform-based correction scheme upon image registration. Notably, by restoring the missing low-frequency content, our hybrid method appears capable of recovering phase values beyond the conventional 0-2π range - an area where conventional FPM techniques often struggle when dealing with optically thick samples. We validated our method using a quantitative phase test target for benchmarking accuracy and biological cheek cells, mouse neurons, and mouse brain tissue slice samples to demonstrate applicability for in vitro bioimaging. Experimental results confirm substantial improvements in phase reconstruction fidelity across spatial frequencies, establishing this hybrid FPM+TIE framework as a practical and high-performance solution for quantitative phase imaging in biomedical and optical metrology applications.

physics.optics

Spatially Multiplexed Interferometric Microscopy with one-dimensional diffraction grating

Digital holographic microscopy (DHM) applied to quantitative phase imaging (QPI) has been successfully demonstrated as a powerful label-free method to analyse the optical properties of cells. Spatially multiplexed interferometric microscopy (SMIM) is a DHM technique that implements a common-path interferometric layout in the embodiment of a standard microscope to achieve QPI. More concretely, SMIM introduces three minimal modifications: 1) replaces the broadband illumination of the microscope by a coherent or partially coherent light source, 2) divides the input plane into two or three regions for transmission in parallel of both imaging and reference beams, and 3) includes a one-dimensional diffraction grating or a beam splitter cube for holographic recording. Hence, SMIM is a cost-effective, extremely simple, and highly stable manner of converting a standard bright field microscope into a holographic one. The goal of this contribution is to provide a review of the SMIM approaches implemented using a one-dimensional (1D) diffraction grating, and highlight vast range of capabilities including super-resolved, reflective, transflective, noise-reduced and single-shot slightly off-axis amplitude and phase imaging.

physics.optics

Variable zoom digital in-line holographic microscopy

We report on a novel layout providing variable zoom in digital in-line holographic microscopy (VZ-DIHM). The implementation is in virtue of an electrically tunable lens (ETL) which enables to slightly shift the illumination source axial position without mechanical movement of any system component. Magnifications ranging from 15X to 35X are easily achievable using the same layout and resulting in a substantial variation of the total field of view (FOV). The performance of the proposed setup is, first, validated using a resolution test target where the main parameters are analyzed (theoretically and experimentally) and, second, corroborated analyzing biological sample (prostate cancer cells) showing its application to biomedical imaging.

physics.optics

Gradient Optical Diffraction Tomography

Optical diffraction tomography (ODT) enables non-invasive information-rich 3D refractive index (RI) reconstruction of unimpaired transparent biological and technical samples, crucial in biomedical research, optical metrology, materials sciences, and other fields. ODT bypasses the inherent limitations of 2D integrated quantitative phase imaging methods. To increase the signal-to-noise ratio easy-to-implement common-path shearing interferometry setups are successfully combined with low spatiotemporal coherence of illumination. The need for self-interference generated holograms, with small shear values, critically impedes the analysis of dense and thick samples, e.g., cell cultures, tissue sections, and embryos/organoids. Phase gradient imaging techniques, deployed as a popular solution in the small shear regime, up to now were constrained to 2D integrated quasi-quantitative phase imaging and z-scanning for depth resolution. To fill this significant scientific gap, we propose a novel gradient optical diffraction tomography (GODT) method. The GODT uses coherence-tailored illumination-scanning sequence of phase gradient measurements to tomographically reconstruct, for the first time to the best of our knowledge, a derivative of the 3D RI distribution (in the shear direction) with clearly visible 3D sample structure and high sensitivity to its spatial variations. We present a mathematically rigorous theory based on the first-order Rytov approximation behind the new method, validate it using simulations deploying numerical Shepp-Logan target and corroborate experimentally via successful tomographic imaging of the calibrated nano-printed cell phantom and efficient examination of neural cells. This novel imaging modality opens new possibilities in biomedical quantitative phase imaging, advancing the field and putting forward a first of a kind contrast domain: 3D RI gradient.

physics.optics