Digital Holographic Imaging Research Articles

Quantitative label-free digital holographic imaging of cardiomyocyte optical volume, nucleation, and cell division

Cardiac regeneration in newborn rodents depends on the ability of pre-existing cardiomyocytes to proliferate and divide. This capacity is lost within the first week of postnatal development when these cells rapidly switch from hyperplasia to hypertrophy, withdraw from the cell cycle, become binucleated, and increase in size. How these dynamic changes in cell size and nucleation impact cardiomyocyte proliferative potential is not well understood. In this study, we innovate the application of a commercially available digital holographic imaging microscope, the Holomonitor M4, to evaluate the proliferative responses of mononucleated and binucleated cardiomyocytes after CHIR99021 treatment, a model proliferative stimulus. This system enables long-term label-free quantitative tracking of primary cardiomyocyte dynamics in real-time with single-cell resolution. Our results confirm that chemical inhibition of glycogen synthase kinase 3 with CHIR99021 promotes complete cell division of both mononucleated and binucleated cardiomyocytes with high frequency. Quantitative tracking of cardiomyocyte volume dynamics during these proliferative events revealed that both mononucleated and binucleated cardiomyocytes reach a similar size-increase threshold prior to attempted cell division. Binucleated cardiomyocytes attempt to divide with lower frequency than mononucleated cardiomyocytes, which may be associated with inadequate increases in cell size. By defining the interrelationship between cardiomyocyte size, nucleation, and cell cycle control, we may better understand the cellular mechanisms that drive the loss of mammalian cardiac regenerative capacity after birth.

Speckle denoising based on Swin-UNet in digital holographic interferometry

Speckle noise, mechano-physical noise, and environmental noise are inevitably introduced in digital holographic coherent imaging, which seriously affects the quality of phase maps, and the removal of non-Gaussian statistical noise represented by speckle noise has been a challenging problem. In the past few years, deep learning methods based on convolutional neural networks (CNNs) have made good progress in removing Gaussian noise. However, they tend to fail when these deep networks designed for Gaussian noise removal are used to remove speckle noise. Recently, numerous studies have employed CNNs to address the issue of degraded speckle images, yielding encouraging results. Nevertheless, the degradation of speckle noise that is simulated in isolation is limited and insufficient to encompass the increasingly complex DHI noise environment. This paper presents what we believe to be a novel approach to simulating complex noise environments by multiplexing simulated Gaussian noise and speckle noise. The noise resulting from aliasing does not adhere to the statistical laws of the noise prior to aliasing, which poses a more challenging task for the noise-reduction algorithms utilized in neural networks. Consequently, in conjunction with the capacity of the Swin Transformer to model multi-scale features, this paper proposes a DHI speckle denoising approach based on Swin-UNet. In this paper, Gaussian, speckle, and blending noise datasets with different noise densities are constructed for training and testing by numerical simulation, and generalizability tests are performed on 1,100 randomly selected open-source holographic tomography (HT) noise images at Warsaw University of Technology and 25 speckle images selected from DATABASE. All test results are quantitatively evaluated by three evaluation metrics: mean squared error (MSE), peak signal-to-noise ratio (PSNR), and structural similarity index (SSIM). All convolutional neural network (CNN) algorithms are evaluated qualitatively based on the number of parameters, floating point operations, and denoising time. The results of the comparison demonstrate that the denoising algorithm presented in this paper exhibits greater stability, accuracy, and generalizability.

Compact common-path polarization holography for measurement of the Jones matrix of polarization-sensitive materials

A compact common-path off-axis digital holographic imaging method is proposed utilizing polarization-angular-multiplexing for Jones matrix measurement. Our method employs a common-path off-axis configuration to capture multiplexed off-axis interferograms generated by orthogonally polarized object beams and a reference beam on a monochrome CCD camera. The modulation of the fringe direction is achieved by two homemade retro-reflector mirrors, allowing for the retrieval of the Jones matrix distribution of transparent specimens through a matrix-division algorithm. The stable common-path design and the expansive camera field of view facilitate the extraction of spatially resolved Jones matrix parameters. The feasibility of this method was validated through experiments involving standard objects and polarization-sensitive materials, conducted at both general and microscopic scales. Our system completed polarization imaging of cancerous tissues, unequivocally demonstrating its extraordinary potential in medical pathology diagnostics.

Speckle noise suppression of a reconstructed image in digital holography based on the BM3D improved convolutional neural network

In digital holographic measurement, when light waves pass through inhomogeneous media or surfaces, speckle noise is generated, resulting in random, granular light and dark spots in the hologram, which greatly reduces the image quality. Therefore, in order to improve the image quality of holographic reconstruction, a noise reduction method based on the BM3D improved convolutional neural network (CNN) is proposed in this paper. Firstly, the similarity and important statistical information between blocks can be obtained by using BM3D. Then, the denoising convolutional neural network (DnCNN) is used to learn the relationship between the noise of a large number of samples and the noise image, and further purify the image to retain the details for a better denoising effect. Finally, a reflective off-axis digital holographic optical path system is constructed to collect the holograms of the test samples, and the reconstructed images are obtained by the Fresnel diffraction method to constitute a dataset with the simulated holographic reconstructed images to validate the proposed method in this paper, compared to the other methods, such as DnCNN, convolutional blind denoising network (CBDNet), BM3D, and Wiener filtering. The experimental results of qualitative and quantitative analyses show that the proposed method combines the advantages of traditional algorithms and deep learning, significantly enhances the robustness of the system, optimizes the denoising performance, and preserves the details of the reconstructed image to the greatest extent.

Range selective digital holographic imaging of vibrating objects using FMCW lidar

The use of frequency modulated continuous wave (FMCW) chirped transmit and reference waveforms in digital holographic (DH) imaging has enabled range selectivity. By frequency shifting the reference beam to compensate for the typical FMCW lidar beat frequency associated with a particular range, a temporally stable holographic image is formed for objects at the selected range and coherently integrates on a short wave infrared (SWIR) sensor. For vibrating objects, longitudinal movements of the object greater than half of an optical wavelength during the exposure time of the sensor array induce phase shifts that can wash out the hologram. An analog feedback system was designed and constructed whereby a lidar subassembly provides real time phase compensation information to a DH subassembly in order to stabilize the range selective digital holographic recording of the object. The design and characterization of the feedback system, as well as the results demonstrating the performance for vibrating objects that move over 17 wavelengths during the sensor exposure, are discussed.

Multi-Perspective Heterodyne Digital Holography For High-Speed Imaging Of Density Fluctuations In Supersonic Flow

Density fluctuation measurements with high spatio-temporal resolution provide meaningful insights into the dynamics of turbulent structures in shear layers, boundary layers, and disturbances that influence the transition to turbulence. The current contribution presents a new multiple perspective digital holographic imaging technique that aims to resolve high-frequency density fluctuations with spatial discrimination along the line of sight. A key element of the proposed method is the generation of a beamlet array of varying view angles that is spatially overlapped within the measurement zone. We present the first experimental realization of this approach using a 7-element hexagonally packed fly’s eye lens array and analyze results for a pair of mutually perpendicular and axially offset sonic jets. The experimental apparatus also incorporates heterodyne detection with a heterodyne frequency set to 1/4 of the imaging frame rate. The optical system performance is evaluated for a crossing jet flow and key design considerations for the multi-perspective optical system are discussed.

O-113 Non-invasive evaluation of embryo metabolic activity through hyperspectral microscopy

Abstract Current approaches for assessing embryo quality are subjective (morphology) or invasive (biopsy) and are not always predictive of live birth. Development of an accurate and non-invasive method to assess embryo quality would likely improve IVF success. We have taken a different approach towards developing a non-invasive diagnostic for embryo quality: optical imaging. We use low levels of light to illuminate the embryo and obtain spatial information on cell metabolism as well as biophysical properties of the embryo. Here I will present our work showing that hyperspectral imaging can be used to accurately discriminate between euploid and aneuploid mouse embryos, and in light sheet mode, able to generate 3D metabolic maps of developing embryos. Importantly, we show that this form of imaging is safe: compared to embryos that were not imaged, hyperspectral imaged embryos had similar, and not significantly different, pregnancy and implantation rates, with offspring weight at weaning also comparable. Another potential optical approach to assess embryo quality is digital holographic imaging. To date, little attention has been paid to the physical parameters of an embryo and whether these could be markers of embryo quality. One such physical parameter is refractive index. We have shown that digital holographic microscopy is able to measure very small changes in refractive index between good and poor-quality mouse embryos. Importantly, these optical approaches occur in the absence of stains or exogenous tags. Used alone, or in combination, such optical approaches may prove beneficial in developing an accurate, non-invasive diagnostic for embryo quality.

Deeper Insights into Mixed Crowding through Enzyme Activity, Dynamics, and Crowder Diffusion.

Given the fact that the cellular interior is crowded by many different kinds of macromolecules, it is important that in vitro studies be carried out in the presence of mixed crowder systems. In this regard, we have used binary crowders formed by the combination of some of the commonly used crowding agents, namely, Ficoll 70, Dextran 70, Dextran 40, and PEG 8000 (PEG 8), to study how these affect enzyme activity, dynamics, and crowder diffusion. The enzyme chosen is AK3L1, an isoform of adenylate kinase. To investigate its dynamics, we have carried out three single point mutations (A74C, A132C, and A209C) with the cysteine residues being labeled with a coumarin-based solvatochromic probe [CPM: (7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin)]. Both enzyme activity and dynamics decreased in the binary mixtures as compared with the sum of the individual crowders, suggesting a reduction in excluded volume (in the mixture). To gain deeper insights into the binary mixtures, fluorescence correlation spectroscopy studies were carried out using fluorescein isothiocyanate-labeled Dextran 70 and tetramethylrhodamine-labeled AK3L1 as the diffusion probes. Diffusion in binary mixtures was observed to be much more constrained (relative to the sum of the individual crowders) for the labeled enzyme as compared to the labeled crowder showing different environments being faced by the two species. This was further confirmed during imaging of the phase-separated droplets formed in the binary mixtures having PEG as one of the crowding agents. The interior of these droplets was found to be rich in crowders and densely packed, as shown by confocal and digital holographic microscopy images, with the enzymes predominantly residing outside these droplets, that is, in the relatively less crowded regions. Taken together, our data provide important insights into various aspects of the simplest form of mixed crowding, that is, composed of just two components, and also hint at the enhanced complexity that the cellular interior presents toward having a detailed and comprehensive understanding of the same.

Visiting experience of digital museum interior design based on virtual reality and sensor networks

China's computer technology and image sensor technology are both rapidly developing, and the related advantages of digital holography technology are gradually expanding. These technologies have been applied in many fields. For example, three-dimensional measurement, deformation measurement, and anti-counterfeiting fields. This article combines holographic technology and virtual reality technology, and applies them to the interior design of digital museums and the optimization of auxiliary visits. A corresponding mathematical calculation model is established for digital holographic images, and in-depth research is conducted on this mathematical calculation model. Relevant personnel have also conducted relevant research on commonly used digital holography technology, and provided a detailed introduction to the principles of virtual reality technology. They have also conducted in-depth research on the specific working principles of VR technology and the application technology of virtual reality systems. Finally, an analysis and research were conducted on the development of Chinese museums, and a new perspective was proposed on the indoor space of museums under the new digital concept, which has certain reference significance. Currently, Chinese museums are thriving, so deepening the interior space of museums is very important. This article hopes to provide some guidance and practical value for the development of Chinese museums.

Fractional Fourier-transform off-axis digital holographic imaging

A fractional Fourier-transform digital holographic imaging method with resolution enhancement features is presented. In an optical configuration, an extended fractional Fourier-transform optical setup is set in the object arm of an off-axis digital holographic recording system to record a fractional Fourier-transform hologram via the optical interference of the fractional Fourier-transform wavefront of an object wave with a reference wave. For reconstruction imaging, the reconstruction approach for fractional Fourier-transform holograms is given. In the experiment, the fractional Fourier-transform digital holograms are recorded under the different recording parameters, and their amplitude images are effectively reconstructed. The imaging results demonstrate that the reconstruction-imaging resolution of fractional-order Fourier-transform holograms is obviously enhanced compared to that of conventional image-plane holograms. The presented fractional Fourier-transform digital holographic imaging with resolution enhancement and optical configuration flexibility provides, to our knowledge, a novel way for off-axis digital holographic imaging.

Application of Digital Holographic Imaging to Monitor Real-Time Cardiomyocyte Hypertrophy Dynamics in Response to Norepinephrine Stimulation.

Cardiomyocyte hypertrophy, characterized by an increase in cell size, is associated with various cardiovascular diseases driven by factors including hypertension, myocardial infarction, and valve dysfunction. In vitro primary cardiomyocyte culture models have yielded numerous insights into the intrinsic and extrinsic mechanisms driving hypertrophic growth. However, due to limitations in current approaches, the dynamics of cardiomyocyte hypertrophic responses remain poorly characterized. In this study, we evaluate the application of the Holomonitor M4 digital holographic imaging microscope to track dynamic changes in cardiomyocyte surface area and volume in response to norepinephrine treatment, a model hypertrophic stimulus. The Holomonitor M4 permits non-invasive, label-free imaging of three-dimensional changes in cell morphology with minimal phototoxicity, thus enabling long-term imaging studies. Untreated and norepinephrine-stimulated primary neonatal rat cardiomyocytes were live-imaged on the Holomonitor M4, which was followed by image segmentation and single-cell tracking using the HOLOMONITOR App Suite software version 4.0.1.546. The 24 h treatment of cultured cardiomyocytes with norepinephrine increased cardiomyocyte spreading and optical volume as expected, validating the reliability of the approach. Single-cell tracking of both cardiomyocyte surface area and three-dimensional optical volume revealed dynamic increases in these parameters throughout the 24 h imaging period, demonstrating the potential of this technology to explore cardiomyocyte hypertrophic responses with greater temporal resolution; however, technological limitations were also observed and should be considered in the experimental design and interpretation of results. Overall, leveraging the unique advantages of the Holomonitor M4 digital holographic imaging system has the potential to empower future work towards understanding the molecular and cellular mechanisms underlying cardiomyocyte hypertrophy with enhanced temporal clarity.

Correction of the wavelength error in transmission of a full-color holographic 3D image

When a digital holographic image represented by a sampled wavefield is transmitted and the wavelength used in the three-dimensional (3D) display devices does not agree exactly with the wavelength of the original image data, the reconstructed 3D image will differ slightly from the original. This slight change is particularly problematic for full-color 3D images reconstructed using three wavelengths. A method is proposed here to correct the holographic image data and reduce the problems caused by wavelength mismatch. The effectiveness of the method is confirmed via theoretical analysis and numerical experiments that evaluate the reconstructed images using several image indices.

HoloDiffusion: Sparse Digital Holographic Reconstruction via Diffusion Modeling

In digital holography, reconstructed image quality can be primarily limited due to the inability of a single small aperture sensor to cover the entire field of a hologram. The use of multi-sensor arrays in synthetic aperture digital holographic imaging technology contributes to overcoming the limitations of sensor coverage by expanding the area for detection. However, imaging accuracy is affected by the gap size between sensors and the resolution of sensors, especially when dealing with a limited number of sensors. An image reconstruction method is proposed that combines physical constraint characteristics of the imaging object with a score-based diffusion model, aiming to enhance the imaging accuracy of digital holography technology with extremely sparse sensor arrays. Prior information of the sample is learned by the neural network in the diffusion model to obtain a score function, which alternately constrains the iterative reconstruction process with the underlying physical model. The results demonstrate that the structural similarity and peak signal-to-noise ratio of the reconstructed images using this method are higher than the traditional method, along with a strong generalization ability.

Digital Holography and 3D Imaging 2023: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

The Optica Topical Meeting on Digital Holography and 3D Imaging (DH) was held 14-17 August 2023 in Boston, Massachusetts. The meeting was organized co-jointly with the Optica Imaging Congress. Feature issues based on the DH meeting series have been released by Applied Optics (AO) since 2007. Since 2017, AO and the Journal of the Optical Society of America A (JOSA A) have presented a feature issue in each journal. This feature issues includes 17 papers in AO and 9 in JOSA A. Together they cover a large range of topics, reflecting the rapidly expanding techniques and applications of digital holography and 3D imaging. The upcoming DH Conference (DH 2024) will be held from 3 to 6 June in Paestum, Italy.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

The Optica Topical Meeting on Digital Holography and 3D Imaging (DH) was held 14-17 August 2023 in Boston, Massachusetts. The meeting was organized co-jointly with the Optica Imaging Congress. Feature issues based on the DH meeting series have been released by Applied Optics (AO) since 2007. Since 2017, AO and the Journal of the Optical Society of America A (JOSA A) have presented a feature issue in each journal. This feature issues includes 17 papers in AO and 9 in JOSA A. Together they cover a large range of topics, reflecting the rapidly expanding techniques and applications of digital holography and 3D imaging. The upcoming DH Conference (DH 2024) will be held from 3 to 6 June in Paestum, Italy.

Unsupervised Deep Learning Enables 3D Imaging for Single‐Shot Incoherent Holography

AbstractIncoherent digital holography no longer requires spatial coherence of the light field, breaking through the imaging resolution of coherent digital holography. However, traditional reconstruction methods cannot avoid the inherent contradiction between temporal resolution and signal‐to‐noise ratio, which is mitigated by deep learning methods, and there are problems such as dataset labeling and insufficient generalization ability. Here, a self‐calibrating reconstruction approach with an untrained network is proposed by fusing the plug‐and‐play nonlinear reconstruction block, the forward physics imaging model, and a physically enhanced neural network. Measurement consistency and total variation kernel function regularization are used to optimize the network parameters and invert the potential process. The results show that the proposed method can achieve high fidelity, high signal‐to‐noise ratio, dynamic, and artifact‐free 3D reconstruction using a single hologram without the need for datasets or labels. In addition, the peak signal‐to‐noise ratio of the reconstructed image with the proposed method is improved by a factor of 4.6 compared to the previous methods. The proposed method leads to considerable performance improvement on the imaging inverse problem, bringing new enlightenment for high‐precision unsupervised incoherent digital holographic 3D imaging.

Hologram Noise Model for Data Augmentation and Deep Learning

This paper introduces a noise augmentation technique designed to enhance the robustness of state-of-the-art (SOTA) deep learning models against degraded image quality, a common challenge in long-term recording systems. Our method, demonstrated through the classification of digital holographic images, utilizes a novel approach to synthesize and apply random colored noise, addressing the typically encountered correlated noise patterns in such images. Empirical results show that our technique not only maintains classification accuracy in high-quality images but also significantly improves it when given noisy inputs without increasing the training time. This advancement demonstrates the potential of our approach for augmenting data for deep learning models to perform effectively in production under varied and suboptimal conditions.

Geometric-Optical Model of Digital Holographic Particle Recording System and Features of Its Application

The paper proposes an equivalent optical scheme of an in-line digital holographic system for particle recording and a mathematical model that establishes a one-to-one correspondence between the dimensional and spatial parameters of a digital holographic image of a particle and the imaged particle itself. The values of the model coefficients used to determine the real size and longitudinal coordinate of a particle according to its holographic image are found by calibration. The model was tested in field and laboratory conditions to calibrate a submersible digital holographic camera designed to study plankton in its habitat. It was shown that four calibration measurements are sufficient enough to determine the model coefficients, and the developed design of the submersible digital holographic camera makes it possible to perform these measurements during the recording of each hologram. In addition, this neither requires data on the refractive index of the medium with particles nor on the parameters of the optical elements of the scheme. The paper presents the results of marine experiments in the Kara Sea and the Laptev Sea, as well as in fresh water in laboratory conditions and in Lake Baikal. The error in measuring the particle size in seawater without the use of the model is 53.8%, while the error in determining their longitudinal coordinates is 79.3%. In fresh water, the same errors were 59% and 54.5%, respectively. The error in determining the position of a particle with the use of the designed mathematical model does not exceed 1.5%, and the error in determining the size is 4.8%. The model is sensitive to changes in the optical properties of the medium, so it is necessary to perform calibration in each water area, and one calibration is quite sufficient within the same water area. At the same time, the developed design of the submersible holographic camera allows, if necessary, calibration at each holographing of the medium volume with particles.

Digital Holographic Particle Field Imaging Based on Infrared Illumination (Invited)

Digital Holographic Particle Field Imaging Based on Infrared Illumination (Invited)

Digital Holographic Focusing Imaging Based on Butterworth Feature Function

Digital Holographic Focusing Imaging Based on Butterworth Feature Function