Imaging Of Biological Tissues Research Articles

Cross-sectional imaging of speed-of-sound distribution using photoacoustic reversal beacons

Photoacoustic tomography (PAT) enables non-invasive cross-sectional imaging of biological tissues, but it fails to map the spatial variation of speed-of-sound (SOS) within tissues. While SOS is intimately linked to density and elastic modulus of tissues, the imaging of SOS distribution serves as a complementary imaging modality to PAT. Moreover, an accurate SOS map can be leveraged to correct for PAT image degradation arising from acoustic heterogeneities. Herein, we propose a method for SOS imaging using scanned photoacoustic beacons excited by short laser pulse with inversion reconstruction. Our method is based on photoacoustic reversal beacons (PRBs), which are small light-absorbing targets with strong photoacoustic contrast. We excite and scan a number of PRBs positioned at the periphery of the target, and the generated photoacoustic waves propagate through the target from various directions, thereby achieve spatial sampling of the internal SOS. By picking up the PRB signal using a graph-based dynamic programing algorithm, we formulate a linear inverse model for pixel-wise SOS reconstruction and solve it with iterative optimization technique. We validate the feasibility of the proposed method through simulations, phantoms, and ex vivo biological tissue tests. Experimental results demonstrate that our approach can achieve accurate reconstruction of SOS distribution. Leveraging the obtained SOS map, we further demonstrate significantly enhanced PAT image reconstruction with acoustic correction.

Exploring the Interplay of Wavelength, Quantum Yield, and Penetration Depth in In Vivo Fluorescence Imaging.

The intricate interplay between the irradiation wavelength, the fluorophore quantum yield (QY) and penetration depth profoundly influences the efficacy of in vivo fluorescence imaging in various applications. Understanding the complex behavior of fluorescence in vivo, specifically how variations in wavelength affect the QY of commonly used dyes and the depth of imaging is crucial for optimizing fluorescence imaging techniques, as it directly impacts the accuracy and efficiency of imaging in biological tissues. In our study, we explore these dynamics through Monte Carlo simulations conducted under conditions reflective of wide-field fluorescence imaging, examining how variations in wavelength impact the dye's QY and depth of imaging, and consequently, the fluorescence behavior. A transition in the exponential decay of the emission depth exponent is observed around the 500-600nm range, indicating varying degrees of influence of depth on the fluorescence emission. The analysis of the fluorophore's QY reveals wavelength-dependent variations, with the most significant impact observed in the 600-700nm range. Moreover, we continued our investigation to explore multiplexing, unveiling insights into the spacing between identical spots in multiplexing images across various depths and wavelengths.

Nonlinear acoustic modulation utilizing designed acoustic bubble array.

Acoustic modulation has attracted significant investigative interest for their outstanding promising application scenes. Furthermore, acoustic bubble array has shown anticipated foreground in signal processing and acoustic manipulation. Here, we demonstrate a nonlinear acoustic modulation method via designed acoustic bubble array. Numerical calculations have been conducted to analyze several influential parameters and the corresponding effects on the vibrational behaviors of the acoustic bubbles. Appropriate corrections have been added on the numerical model to elucidate the physical scene. Experimental validation has confirmed the practicability and validity of the designation. Potential applications in biological tissue imaging and unidirectional sound transmission can be expected with further research.

Evaluating the effects of three anesthetic regimens on retinal structure and function in the living mouse eye using a customized bimodal OCT/ORG system

Abstract Optical coherence tomography (OCT) is a non-invasive imaging technique that generates cross-sectional images of biological tissues at millimeter-level penetration depth with micron-level resolution. In this study, we developed a custom spectral-domain OCT system equipped with a flash stimulus module for dual-modality imaging of mouse retinal structure and function. The axial resolution of the system reached ∼2.7/2 μm in air/tissue following optimization and calibration. We deployed this system to conduct in vivo OCT structural imaging and optoretinography (ORG) functional assessment of mouse retina to investigate the effects of three anesthesia regimens. Statistical analysis of the results indicates that anesthesia results in a smaller relative intensity of inner/outer segment junction and external limiting membrane and a thickening of the outer segment compared to an awake state. Meanwhile, significant differences were observed in the effects on retinal structure and function between isoflurane and tribromoethanol anesthesia, while differences between isoflurane and esketamine+xylazine groups were small. These findings suggest that the type of anesthetic should be taken into account when comparing imaging results across studies reported in the literature.

Magnetic Induction Tomography for Brain Tissue Imaging Based on Conductivity Distribution for Parkinson’s Disease Diagnosis

Parkinson's disease is a prevalent neurodegenerative complication defined by the accumulation of alpha synuclein lewy bodies in the brain. Misdiagnosis results widespread of Parkinson’s disease because clinical diagnosis is challenging, underlining a need of a better detection technique, such as non-invasive magnetic induction tomography (MIT) technique. Non-invasive techniques for biological tissues imaging are becoming popular in biomedical engineering field. Therefore, MIT technology as a non-invasive technique has been encouraged in a medical field due to its advancement of technology in diagnosing diseases. The measurement parameters in MIT are passive electromagnetic properties (conductivity, permittivity, permeability) for biological tissue and the most dominant parameter in MIT is conductivity properties. It is uses a phase shift between a primary magnetic field and an induced field caused by a target object's conductivity. As a function of conductivity, the phase shift between the applied and secondary fields is expressed. Thus, the phase shift can be used to characterize the conductivity of a target object. The phase shift between the excitation and induced magnetic fields (EMF and IMF) reflects the change in conductivity in biological tissues. This paper focuses on the virtual simulation by using COMSOL Multi-physics for the design and development of MIT system that emphasizes on single channel magnetic induction tomography for biological tissue (bran tissue) imaging based on conductivity distribution for Parkinson’s disease diagnosis. The develop system employs the use of excitation coils to induce an electromagnetic field (e.m.f) in the brain tissue, which is then measured at the receiving side by sensors. The proposed system is capable of indicating Parkinson’s disease based on conductivity distribution. This method provides the valuable information of the brain abnormality based on differences of conductivities of normal brain and Parkinson’s disease brain tissues.

Synchrotron X-ray imaging of soft biological tissues - principles, applications and future prospects.

Synchrotron-based tomographic phase-contrast X-ray imaging (SRµCT or SRnCT) is a versatile isotropic three-dimensional imaging technique that can be used to study biological samples spanning from single cells to human-sized specimens. SRµCT and SRnCT take advantage of the highly brilliant and coherent X-rays produced by a synchrotron light source. This enables fast data acquisition and enhanced image contrast for soft biological samples owing to the exploitation of phase contrast. In this Review, we provide an overview of the basics behind the technique, discuss its applications for biologists and provide an outlook on the future of this emerging technique for biology. We introduce the latest advances in the field, such as whole human organs imaged with micron resolution, using X-rays as a tool for virtual histology and resolving neuronal connections in the brain.

Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging

The ultimate aim of fluorescence microscopy is to achieve high-resolution imaging of increasingly larger biological samples. Extended depth of field presents a potential solution to accelerate imaging of large samples when compression of information along the optical axis is not detrimental to the interpretation of images. We have implemented an extended depth of field (EDF) approach in a random illumination microscope (RIM). RIM uses multiple speckled illuminations and variance data processing to double the resolution. It is particularly adapted to the imaging of thick samples as it does not require the knowledge of illumination patterns. We demonstrate highly-resolved projective images of biological tissues and cells. Compared to a sequential scan of the imaged volume with conventional 2D-RIM, EDF-RIM allows an order of magnitude improvement in speed and light dose reduction, with comparable resolution. As the axial information is lost in an EDF modality, we propose a method to retrieve the sample topography for samples that are organized in cell sheets.

Non-Invasive Bio-Impedance Imaging and Sensing for Medical Diagnostics and Industrial Applications

Bio-impedance-based imaging technique stands at the intersection of medical/industrial imaging techniques and is also known as electrical impedance tomography (EIT), imparting a non-invasive, value-powerful approach for imaging biological tissues and procedures. We begin by elucidating the essential standards underlying EIT, focusing on the size of impedance variations inside an object (plastic pipe) to reconstruct its inner conductivity distribution. Subsequently, we delve into the numerous programs of EIT across numerous fields and discuss recent experimental strategies and advancements aimed at mitigating these boundaries, such as novel electrode configurations, advanced reconstruction algorithms, and incorporation of complementary imaging modalities. Finally, we underscore the importance of experimental research in advancing the capabilities and packages of EIT as a valuable device in industrial monitoring, specifically in bedside tracking, process pipelines, tank monitoring and also used for other medical or industrial detection, even as additionally exploring its rising roles in commercial and environmental applications.

Broadband SWIR emission through Cr3+-Ni2+ energy transfer in YAGG fluorescent ceramics for bioimaging applications

Short-wave infrared (SWIR) light in the 1000–1700 nm range has high penetration capability through biological tissues and is non-destructive, making it widely used in biomedical applications and near-infrared imaging. Currently, the development of SWIR light sources is urgently needed. This paper discusses the luminescent properties of the near-infrared emitting transition metal ions Cr3+ and Ni2+. By using the traditional high-temperature solid-state method, we successfully synthesized YAGG:3.5 %Cr3+-0.5 %Ni2+ (YAGG:Cr-Ni) garnet-based phosphor ceramic in one step. The phosphor ceramic emits ultra-broadband NIR-I (600–900 nm, FWHM=131 nm) and SWIR (1100–1700 nm, FWHM=377 nm) light when excited by a 450 nm commercial blue LED. Position analysis combined with density functional theory (DFT) simulation confirmed the effective substitution of luminescent center ions for the [Al/GaO6] sites. Spectral and lifetime analyses further verified the energy transfer process between Cr3+ and Ni2+ is dipole-quadrupole interaction. Additionally, this phosphor ceramic has a high thermal conductivity of 8.018 W·m⁻¹·K⁻¹ at room temperature. Finally, SWIR pc-LED devices were fabricated by packaging the phosphor ceramic with a 450 nm commercial blue LED, achieving skeletal and venous imaging of biological tissues up to 20 mm thick, providing a new approach for the development of SWIR devices.

Frontiers in artificial intelligence-directed light-sheet microscopy for uncovering biological phenomena and multi-organ imaging.

Light-sheet fluorescence microscopy (LSFM) introduces fast scanning of biological phenomena with deep photon penetration and minimal phototoxicity. This advancement represents a significant shift in 3-D imaging of large-scale biological tissues and 4-D (space + time) imaging of small live animals. The large data associated with LSFM requires efficient imaging acquisition and analysis with the use of artificial intelligence (AI)/machine learning (ML) algorithms. To this end, AI/ML-directed LSFM is an emerging area for multi-organ imaging and tumor diagnostics. This review will present the development of LSFM and highlight various LSFM configurations and designs for multi-scale imaging. Optical clearance techniques will be compared for effective reduction in light scattering and optimal deep-tissue imaging. This review will further depict a diverse range of research and translational applications, from small live organisms to multi-organ imaging to tumor diagnosis. In addition, this review will address AI/ML-directed imaging reconstruction, including the application of convolutional neural networks (CNNs) and generative adversarial networks (GANs). In summary, the advancements of LSFM have enabled effective and efficient post-imaging reconstruction and data analyses, underscoring LSFM's contribution to advancing fundamental and translational research.

High-resolution in vivo 4D-OCT fish-eye imaging using 3D-UNet with multi-level residue decoder.

Optical coherence tomography (OCT) allows high-resolution volumetric imaging of biological tissues in vivo. However, 3D-image acquisition often suffers from motion artifacts due to slow frame rates and involuntary and physiological movements of living tissue. To solve these issues, we implement a real-time 4D-OCT system capable of reconstructing near-distortion-free volumetric images based on a deep learning-based reconstruction algorithm. The system initially collects undersampled volumetric images at a high speed and then upsamples the images in real-time by a convolutional neural network (CNN) that generates high-frequency features using a deep learning algorithm. We compare and analyze both dual-2D- and 3D-UNet-based networks for the OCT 3D high-resolution image reconstruction. We refine the network architecture by incorporating multi-level information to accelerate convergence and improve accuracy. The network is optimized by utilizing the 16-bit floating-point precision for network parameters to conserve GPU memory and enhance efficiency. The result shows that the refined and optimized 3D-network is capable of retrieving the tissue structure more precisely and enable real-time 4D-OCT imaging at a rate greater than 10 Hz with a root mean square error (RMSE) of ∼0.03.

Expanding super-resolution imaging versatility in organisms with multi-confocal image scanning microscopy

Abstract Resolving complex three-dimensional subcellular dynamics noninvasively in live tissues demands imaging tools that balance spatiotemporal resolution, field-of-view and phototoxicity. Image scanning microscopy (ISM), as an advancement of confocal laser scanning microscopy, provides a two-fold 3D resolution enhancement. Nevertheless, the relatively low imaging speed has been the major obstacle for ISM to be further employed in in vivo imaging of biological tissues. Our proposed solution, multi-confocal image scanning microscopy (MC-ISM), aims to overcome the limitations of existing techniques in terms of spatiotemporal resolution balancing by optimizing pinhole diameter and pitch, eliminating out-of-focus signals, and introducing a frame reduction reconstruction algorithm. The imaging speed is increased by 16 times compared with multifocal structured illumination microscopy. We further propose single-galvo scan, akin to the Archimedes spiral in spinning disk confocal system, to ensure high speed ang high accuracy scan without galvanometer's inertial motion. Benefitting from its high photon efficiency, MC-ISM allows continuous imaging of mitochondria dynamics in live cell for 1000 frames without apparent phototoxicity, reaching an imaging depth of 175 μm. Noteworthy, MC-ISM enables the observation of the inner membrane structure of living mitochondria in Arabidopsis hypocotyl for the first time, demonstrating its outstanding performance.

High-Stability Near-Infrared Luminescent Glass Ceramic and Its Applications.

Near-infrared (NIR) light, valuable for its biological penetration and invisibility to the human eye, is a crucial tool in biomedicine, environmental monitoring, anticounterfeiting, and information encryption, yet traditional NIR luminescent materials are often unstable in humid conditions. Here, a highly stable MgGeO3:Mn2+ glass ceramic (GC) with NIR luminescence was successfully synthesized. As-obtained GC700 boasts exceptional luminescent capabilities and possesses abundant trap structures, enabling data inscription with a 405 nm laser and retrieval via laser/thermal excitation. Moreover, the emission peak of Mn2+ can be manipulated from 630 to 691 nm by increasing the annealing treatment temperature. With the harnessing of the effective NIR emission, stable carrier characteristics, and numerous trap structures, there is potential for application in information encryption. Accordingly, we explored the application of MgGeO3:Mn2+ GC (GC700 and GC800) samples in precious three-dimensional (3D) information storage and NIR mechanoluminescence (ML) for biological tissue imaging. These applications demonstrate the potential and versatility of electron-capturing NIR luminescent materials in a range of cutting-edge fields.

Event-responsive scanning transmission electron microscopy.

An ever-present limitation of transmission electron microscopy is the damage caused by high-energy electrons interacting with any sample. By reconsidering the fundamentals of imaging, we demonstrate an event-responsive approach to electron microscopy that delivers more information about the sample for a given beam current. Measuring the time to achieve an electron count threshold rather than waiting a predefined constant time improves the information obtained per electron. The microscope was made to respond to these events by blanking the beam, thus reducing the overall dose required. This approach automatically apportions dose to achieve a given signal-to-noise ratio in each pixel, eliminating excess dose that is associated with diminishing returns of information. We demonstrate the wide applicability of our approach to beam-sensitive materials by imaging biological tissue and zeolite.

Virtual-point-based deconvolution for optical-resolution photoacoustic microscopy.

Optical-resolution photoacoustic microscopy (OR-PAM) has been increasingly utilized for in vivo imaging of biological tissues, offering structural, functional, and molecular information. In OR-PAM, it is often necessary to make a trade-off between imaging depth, lateral resolution, field of view, and imaging speed. To improve the lateral resolution without sacrificing other performance metrics, we developed a virtual-point-based deconvolution algorithm for OR-PAM (VP-PAM). VP-PAM has achieved a resolution improvement ranging from 43% to 62.5% on a single-line target. In addition, it has outperformed Richardson-Lucy deconvolution with 15 iterations in both structural similarity index and peak signal-to-noise ratio on an OR-PAM image of mouse brain vasculature. When applied to an in vivo glass frog image obtained by a deep-penetrating OR-PAM system with compromised lateral resolution, VP-PAM yielded enhanced resolution and contrast with better-resolved microvessels.

Phase-contrast visualization of human tissues using superimposed wavefront imaging of diffraction-enhanced x-rays.

Phase-contrast computed tomography (CT) using high-brilliance, synchrotron-generated x-rays enable three-dimensional (3D) visualization of microanatomical structures within biological specimens, offering exceptionally high-contrast images of soft tissues. Traditional methods for phase-contrast CT; however, necessitate a gap between the subject and the x-ray camera, compromising spatial resolution due to penumbral blurring. Our newly developed technique, Superimposed Wavefront Imaging of Diffraction-enhanced x-rays (SWIDeX), leverages a Laue-case Si angle analyzer affixed to a scintillator to convert x-rays to visible light, capturing second-order differential phase contrast images and effectively eliminating the distance to the x-ray camera. This innovation achieves superior spatial resolution over conventional methods. In this paper, the imaging principle and CT reconstruction algorithm based on SWIDeX are presented in detail and compared with conventional analyzer-based imaging (ABI). It also shows the physical setup of SWIDeX that provides the resolution preserving second-order differential images for reconstruction. We compare the spatial resolution and the sensitivity of SWIDeX to conventional ABI. To demonstrate high-spatial resolution achievable by SWIDeX, the internal structures of four human tissues-ductal carcinoma in situ, normal stomach, normal pancreas, and intraductal papillary mucinous neoplasm of the pancreas-were visualized using an imaging system configured at the Photon Factory's BL14B beamline under the High Energy Accelerator Research Organization (KEK). Each tissue was thinly sliced after imaging, stained with hematoxylin and eosin (H&E) for conventional microscope-based pathology. A comparison of SWIDeX-CT and pathological images visually demonstrates the effectiveness of SWIDeX-CT for biological tissue imaging. SWIDeX could generate clearer 3D images than existing analyzer-based phase-contrast methods and accurately delineate tissue structures, as validated against histopathological images. SWIDeX can visualize important 3D structures in biological soft tissue with high spatial resolution and can be an important tool for providing information between the disparate scales of clinical and pathological imaging.

Navigating challenges and solutions in quantitative photoacoustic imaging

Photoacoustic imaging, an emerging modality that seamlessly combines advantages of optical absorption contrast and ultrasound resolution, holds great promise for noninvasive imaging of biological tissues. Its applications span across diverse fields, such as dermatology, oncology, cardiology, and neurology. However, achieving accurate image reconstruction and physiological parameters quantification from raw photoacoustic signals presents a significant challenge. This challenge primarily arises from the inherent heterogeneity of tissues, encompassing variations in optical fluence and acoustic properties. In addition, incomplete information acquired from a limited view also leads to artifacts, image distortions, and reduced spatial resolution. Furthermore, robust spectral unmixing approach is another key step to restore the initial biochemical components' distribution with complex or unknown background absorption. To overcome these hurdles, researchers have proposed numerous state-of-the-art techniques, aiming to improve the accuracy and reliability of quantitative photoacoustic imaging (qPAI) in heterogeneous tissue. This review aims to comprehensively overview recent developments over the past decade, for addressing four main challenges frequently encountered in qPAI: limited-view reconstruction, acoustic heterogeneity, optical fluence fluctuations, and robust spectral unmixing, which serves as a reference for readers seeking to understand the specific challenges and corresponding solutions in this field.

Non-contact elasticity contrast imaging using photon counting.

Tissues' biomechanical properties, such as elasticity, are related to tissue health. Optical coherence elastography produces images of tissues based on their elasticity, but its performance is constrained by the laser power used, working distance, and excitation methods. We develop a new method to reconstruct the elasticity contrast image over a long working distance, with only low-intensity illumination, and by non-contact acoustic wave excitation. We combine single-photon vibrometry and quantum parametric mode sorting (QPMS) to measure the oscillating backscattered signals at a single-photon level and derive the phantoms' relative elasticity. We test our system on tissue-mimicking phantoms consisting of contrast sections with different concentrations and thus stiffness. Our results show that as the driving acoustic frequency is swept, the phantoms' vibrational responses are mapped onto the photon-counting histograms from which their mechanical properties-including elasticity-can be derived. Through lateral and longitudinal laser scanning at a fixed frequency, a contrast image based on samples' elasticity can be reliably reconstructed upon photon level signals. We demonstrated the reliability of QPMS-based elasticity contrast imaging of agar phantoms in a long working distance, low-intensity environment. This technique has the potential for in-depth images of real biological tissue and provides a new approach to elastography research and applications.

Construction of multifunctional core-shell structure nanoprobe integrating NIR dual-mode optical thermometer and nuclear magnetic imaging

Precise temperature regulation and imaging are critical for in vivo photothermal treatment. The goal of this work is to realize non-contact temperature measurement and imaging in biological tissues by building a multifunctional nano-platform using optical temperature probes and imaging for biological diagnosis and therapy. Here, NIR dual-mode temperature probes of NaYF4: Nd/Yb/Gd@NaNdF4 were designed according to the LIR-I of 4F3/2 → 4I9/2/2F5/2 → 2F7/2 levels in 850 – 1025 nm and LIR-II of 2F5/2 → 2F7/2/4F3/2 → 4I11/2 in 950 – 1100 nm from the emissions of Nd3+/Yb3+. Transmission electron microscope (TEM) and X-ray diffraction (XRD) data of the sample demonstrate its superior dispersibility and accurate structure. The integrated intensity ratio centered around various luminescence intensity peaks is used as a temperature sensor since luminescence intensity is temperature-dependent. Additionally, the temperature sensing capabilities of two ratio temperature sensors were examined under 808 nm laser excitation, utilizing LIR (luminescence intensity ratio) technology. Furthermore, the MRI test indicated that the sample exhibited good imaging capacity when Gd3+ was added. These results assure the auspicious application prospects of the samples in temperature sensing, photothermal therapy, and biomedical imaging.

EDTP enhances and protects the fluorescent signal of GFP in cleared and expanded tissues

Advanced 3D high-resolution imaging techniques are essential for investigating biological challenges, such as neural circuit analysis and tumor microenvironment in intact tissues. However, the fluorescence signal emitted by endogenous fluorescent proteins in cleared or expanded biological samples gradually diminishes with repeated irradiation and prolonged imaging, compromising its ability to accurately depict the underlying scientific problem. We have developed a strategy to preserve fluorescence in cleared and expanded tissue samples during prolonged high-resolution three-dimensional imaging. We evaluated various compounds at different concentrations to determine their ability to enhance fluorescence intensity and resistance to photobleaching while maintaining the structural integrity of the tissue. Specifically, we investigated the impact of EDTP utilization on GFP, as it has been observed to significantly improve fluorescence intensity, resistance to photobleaching, and maintain fluorescence during extended room temperature storage. This breakthrough will facilitate extended hydrophilic and hydrogel-based clearing and expansion methods for achieving long-term high-resolution 3D imaging of cleared biological tissues by effectively safeguarding fluorescent proteins within the tissue.