Imaging Of Biological Tissues Research Articles

Development of organic semiconducting materials for deep-tissue optical imaging, phototherapy and photoactivation.

Biophotonics as a highly interdisciplinary frontier often requires the assistance of optical agents to control the light pathways in cells, tissues and living organisms for specific biomedical applications. Organic semiconducting materials (OSMs) composed of π-conjugated building blocks as the optically active components have recently emerged as a promising category of biophotonic agents. OSMs possess common features including excellent optical properties, good photostability and biologically benign composition. This review summarizes the recent progress in the development of OSMs based on small-molecule fluorophores, aggregation-induced emission (AIE) dyes and semiconducting oligomer/polymer nanoparticles (SONs/SPNs) for advanced biophotonic applications. OSMs have been exploited as imaging agents to transduce biomolecular interactions into second near-infrared fluorescence, chemiluminescence, afterglow or photoacoustic signals, enabling deep-tissue ultrasensitive imaging of biological tissues, disease biomarkers and physiological indexes. By fine-tuning the molecular structures, OSMs can also convert light energy into cytotoxic free radicals or heat, allowing for effective cancer phototherapy. Due to their instant light response and efficient light-harvesting properties, precise regulation of biological activities using OSMs as remote transducers has been demonstrated for protein ion channels, gene transcription and protein activation. In addition to highlighting OSMs as a multifunctional platform for a wide range of biomedical applications, current challenges and perspectives of OSMs in biophotonics are discussed.

Терагерцовый микроскоп на основе эффекта твердотельной иммерсии для визуализации биологических тканей-=SUP=-*-=/SUP=-

A novel method of terahertz (THz) microscopy was proposed for imaging of biological tissues with sub-wavelength spatial resolution. It allows for overcoming the Abbe diffraction limit and provides a sub-wavelength resolution thanks to the solid immersion effect – i.e. to the reduction in the dimensions of electromagnetic beam caustic, when the beam is focused in free space, at a small distance (smaller than the wavelength) behind the medium featuring high refractive index. An experimental setup realizing the proposed method was developed. It uses a backward wave oscillator, as a THz-wave emitter, and a Golay cell, as a THz-wave detector. In this setup, the radiation is focused behind the silicon hemisphere in order to realize the solid immersion effect. The spatial resolution of 0.15λ was demonstrated for the developed microscope, while the measurements were carried out at the wavelength of λ=500 μm, with the metal-air interface as a test object. Such a high spatial resolution represents a significant advantage over that of the previously reported arrangements of solid immersion microscopes. The solid immersion microscopy does not imply using any diaphragms or other near-field probes for achieving the sub-wavelength spatial resolution; thus, it eliminates the energy losses associated with such elements. The proposed methods were applied for imaging of biological tissues, and the observed results highlight its potential in biology and medicine.

Система Фур'є-стокс-поляриметрії полікристалічних мереж біологічних тканин

This article discusses the possibility of using Fourier-Stokes-polarimetry to study the structure of polycrystalline networks of biological tissues. The interrelation between the set of statistical moments of the 1st - 4th order, characterizing the coordinate distribution of the azimuth and ellipticity of the Fourier spectra of polarization-inhomogeneous images of biological tissues and the optical anisotropy parameters of polycrystalline networks, is demonstrated.

A Novel Algorithm for High-Resolution Magnetic Induction Tomography Based on Stacked Auto-Encoder for Biological Tissue Imaging

Magnetic induction tomography (MIT) is a non-invasive and non-contact imaging method that uses an excitation coil to generate time-varying magnetic fields in space and reconstruct the internal conductivity distribution based on the phase difference. In this study, a new MIT reconstruction algorithm was proposed for non-contact measurement and monitoring of the location of the anomaly in the biomedical object of interest. To reconstruct the distribution of electrical characteristics inside the biological tissue, this technique uses a stacked auto-encoder (SAE) neural network composed of a multi-layer automatic encoder. The location and reconstruction accuracy of the anomaly based on SAE and back-projection were compared, and a hemorrhagic stroke was simulated to verify the practicability of the proposed algorithm. The results showed that the relative error of reconstruction based on the SAE network algorithm reached 0.29%, which improved anomaly reconstruction accuracy and reduced the prediction time to 0.02 s. At the same time, the network was used for the reconstruction of hemorrhagic stroke in different locations, amounts, and shapes. Accordingly, the SAE neural network reconstruction algorithm proposed in this study, which can autonomously learn the non-linear relationship between input and output, can solve the defects of the traditional reconstruction algorithm, such as serious artifacts and complex calculations.

Optimization of frequency-doubled Er-doped fiber laser for miniature multiphoton endoscopy.

Frequency-doubled femtosecond Er-doped fiber laser is a low-cost and portable excitation source suitable for multiphoton endoscopy. The frequency-doubled wavelength at 780nm is used to excite the intrinsic fluorescence signal. The frequency-doubling with a periodically poled MgO : LiNbO3 (PPLN) is integrated in the distal end of the imaging head to achieve fiber connection. The imaging speed is further improved by optimizing the excitation laser source. A 0.3-mm length of PPLN crystal is selected and the Er-doped fiber laser is manipulated to match its bandwidth with the acceptance bandwidth of the PPLN. Through this optimization, a reduced pulsewidth of 80fs of the frequency-doubled pulse is achieved. All-fiber dispersion compensation and pulse compression by single mode fiber is conducted, which makes the fiber laser directly fiber-coupled to the imaging head. An imaging speed of 4 frames / s is demonstrated on ex vivo imaging of unstained biological tissues, which is 10 times faster than our previous study using a 1-mm-long PPLN. The results show that miniature multiphoton endoscopy using frequency-doubled Er-doped fiber laser has great potential for clinical applications.

Plasmon-tunable Au@Ag core-shell spiky nanoparticles for surface-enhanced Raman scattering

A facile synthetic approach has been developed to prepare uniform and size-tunable spiky Au@Ag core-shell nanoparticles (NPs) to tailor the localized surface plasmon resonance (LSPR) properties. The gradual assembly of small Au nanocrystals allows the size of spiky Au NPs to be modulated from tens to several hundreds of nanometers by tuning the concentration of initial Au seeds and Au source; and the thickness of the Ag shell can be adjusted with stepwise reduction of Ag(I) ions. The LSPR bands of such spiky Au@Ag core-shell NPs resemble those of pure spiky Au NP cores of similar sizes in near-infrared region, and increasing the Ag shell thickness results in a blue shift and broadening of the LSPR band in the near-infrared region. Additionally, the spiky Au@Ag core-shell NPs exhibit improved surface-enhanced Raman scattering (SERS) activity as compared to the bare spiky Au NPs and spherical Ag@Au NPs. This work has offered a facile route to synthesize plasmonic metal NPs with LSPR band in 650 to 800 nm that show strong enhancement of localized electromagnetic field, which provides an effective SERS substrate for SERS imaging and detection in biological fluids and tissues.

MALDI MS Imaging at Acquisition Rates Exceeding 100 Pixels per Second.

The practicality of matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) applied to molecular imaging of biological tissues is limited by the analysis speed. Typically, a relatively low speed of stop-and-go micromotion of XY stages is considered as a factor substantially reducing the rate with which fresh sample material can be supplied to the laser spot. The sample scan rate in our laboratory-built high-throughput imaging TOF mass spectrometer was significantly improved through the use of a galvanometer-based optical scanner performing fast laser spot repositioning on a target plate. The optical system incorporated into the ion source of our MALDI TOF mass spectrometer allowed focusing the laser beam via a modified grid into a 10-μm round spot. This permitted the acquisition of high-resolution MS images with a well-defined pixel size at acquisition rates exceeding 100pixel/s. The influence of selected parameters on the total MS imaging time is discussed. The new scanning technique was employed to display the distribution of an antitumor agent in 3D colorectal adenocarcinoma cell aggregates; a single MS image comprising 100 × 100pixels with 10-μm lateral resolution was recorded in approximately 70s. Graphical Abstract.

Mueller polarimetric imaging of biological tissues: classification in a decision-theoretic framework.

Mueller polarimetry is increasingly recognized as a powerful modality in biomedical imaging. Nevertheless, principled statistical analysis procedures are still lacking in this field. This paper presents a complete pipeline for polarimetric bioimages, with an application to ex vivo cervical precancer detection. In the preprocessing stage, we evaluate the replacement of pixels by superpixels. In the analysis stage, we resort to decision theory to select and tune a classifier. Performances of the retained classifier are evaluated. Decision theory provides a rigorous and versatile framework, allowing generalization to other pathologies, to other imaging procedures, and to classification problems involving more than two classes.

Imaging acoustic sources through scattering media by using a correlation full-matrix filter

In the inhomogeneous medium, acoustic scattering is always a fundamental challenge for photoacoustic imaging. We implement a correlation full-matrix filter (CFMF) combing with a time reversal operator to improve the imaging quality of acoustic sources in complex media. The correlation full-matrix filtering process extracts the direct wave component from the detected signal and preserve all the useful information at the same time. A location factor is considered in the time reversal operator to compensate for the image distortion and false contrast caused by the limited-view detection. The numerical simulations demonstrate that the proposed approach can perform good imaging quality with the higher image signal-noise ratio and better resolution in an acoustic scattering environment. This scheme might be applied to improve the photoacoustic imaging for inhomogeneous biological tissues.

A compact high-speed full-field optical coherence microscope for high-resolution in vivo skin imaging.

A compact high-speed full-field optical coherence microscope has been developed for high-resolution in vivo imaging of biological tissues. The interferometer, in the Linnik configuration, has a size of 11 × 11 × 5 cm3 and a weight of 210 g. Full-field illumination with low-coherence light is achieved with a high-brightness broadband light-emitting diode. High-speed full-field detection is achieved by using part of the image sensor of a high-dynamic range CMOS camera. En face tomographic images are acquired at a rate of 50 Hz, with an integration time of 0.9 ms. The image spatial resolution is 0.9 μm × 1.2 μm (axial × transverse), over a field of view of 245 × 245 μm2 . Images of human skin, revealing in-depth cellular-level structures, were obtained in vivo and in real-time without the need for stabilization of the subject. The system can image larger fields, up to 1 × 1 mm2 , but at a reduced depth.

Low-cost compact multispectral spatial frequency domain imaging prototype for tissue characterization.

We present a low-cost, compact, and multispectral spatial frequency domain imaging prototype. Illumination components, including 9 LEDs (660 nm - 950 nm) placed on a custom-designed printed circuit board, linear and rotational motors, a printed sinusoidal pattern, and collimation and projection optics as well as the detection components are incorporated in a compact custom-designed 3D-printed probe. Reconstruction of absorption and reduced scattering coefficients is evaluated via imaging tissue mimicking phantoms and potentials of the probe for biological tissue imaging are evaluated via imaging human ovarian tissue ex vivo.

Oxygen-induced leakage of spin polarization in Overhauser-enhanced magnetic resonance imaging: Application for oximetry in tumors

Overhauser-enhanced Magnetic Resonance Imaging (OMRI) is a double resonance technique applied for oxygen imaging in aqueous samples and biological tissues. In this report, we present an improved OMRI approach of oxygen measurement using the single line “Finland” trityl spin probe. Compared to a traditional approach, we introduced an additional mechanism of leakage of spin polarization due to an interaction of a spin system with oxygen. The experimental comparison of the new approach with an oxygen-dependent leakage factor to a traditional approach performed in phantom samples in vitro, and mouse tumor model in vivo, shows improved accuracy of determination of oxygen and contrast agent concentrations.

Highly stable wide-field common path digital holographic microscope based on a Fresnel biprism interferometer

Quantitative phase imaging (QPI) of biological cells and tissues is an important technique useful in the determination of many biophysical parameters such as morphology, refractive index, thickness, cell dry mass, hemoglobin concentration, etc. Off-axis digital holography has been ideal for the QPI of microscopic specimens, but it has lower temporal stability compared to on-axis and common path digital holographic microscopes, which offer higher temporal stability. In this paper, we present a very simple, easy to align yet highly stable common path digital holographic microscope based on a Fresnel biprism interferometer. The system uses a biprism to divide the incoming beam into reference and object beams without the loss of optical power, unlike diffraction phase microscopy. Two methods are proposed, one by utilizing the spatial filtering mode and the other by the self-referencing mode. It offers the advantage that the reference beam can be easily created simply by translating the object in the focal plane of the microscopic objective or by spatially filtering one of the object beams in the Fourier domain. The proposed setup offers no power loss and a high phase stability of approximately 0.006 rad (6 mrad) without using any vibration isolation. The experiments on industrial and biological samples are reported demonstrating its application both for static and dynamic samples.

Reflection-mode continuous-wave 0.15λ-resolution terahertz solid immersion microscopy of soft biological tissues

We have developed a method of terahertz (THz) solid immersion (SI) microscopy for continuous-wave reflection-mode imaging of soft biological tissues with a sub-wavelength spatial resolution. In order to achieve strong reduction in the dimensions of the THz beam caustic, an electromagnetic wave is focused into the evanescent field volume behind a medium with a high refractive index. We have experimentally demonstrated a 0.15λ-resolution of the proposed imaging modality at λ = 500 μm, which is beyond the Abbe diffraction limit and represents a considerable improvement over the previously-reported arrangements of SI imaging setups. The proposed technique does not involve any sub-wavelength near-field probes and diaphragms, thus, avoiding the THz beam attenuation due to such elements. We have applied the developed method for THz imaging of various soft tissues: a plant leaf blade, cell spheroids, and tissues of the breast ex vivo. Our THz images clearly reveal sub-wavelength features in tissues, therefore, promising applications of THz SI microscopy in biology and medicine.

Near-Infrared Photoactivatable Nitric Oxide Donors with Integrated Photoacoustic Monitoring.

Photoacoustic (PA) tomography is a noninvasive technology that utilizes near-infrared (NIR) excitation and ultrasonic detection to image biological tissue at centimeter depths. While several activatable small-molecule PA sensors have been developed for various analytes, the use of PA molecules for deep-tissue analyte delivery and monitoring remains an underexplored area of research. Herein, we describe the synthesis, characterization, and in vivo validation of photoNOD-1 and photoNOD-2, the first organic, NIR-photocontrolled nitric oxide (NO) donors that incorporate a PA readout of analyte release. These molecules consist of an aza-BODIPY dye appended with an aryl N-nitrosamine NO-donating moiety. The photoNODs exhibit chemostability to various biological stimuli, including redox-active metals and CYP450 enzymes, and demonstrate negligible cytotoxicity in the absence of irradiation. Upon single-photon NIR irradiation, photoNOD-1 and photoNOD-2 release NO as well as rNOD-1 or rNOD-2, PA-active products that enable ratiometric monitoring of NO release. Our in vitro studies show that, upon irradiation, photoNOD-1 and photoNOD-2 exhibit 46.6-fold and 21.5-fold ratiometric turn-ons, respectively. Moreover, unlike existing NIR NO donors, the photoNODs do not require encapsulation or multiphoton activation for use in live animals. In this study, we use PA tomography to monitor the local, irradiation-dependent release of NO from photoNOD-1 and photoNOD-2 in mice after subcutaneous treatment. In addition, we use a murine model for breast cancer to show that photoNOD-1 can selectively affect tumor growth rates in the presence of NIR light stimulation following systemic administration.

Volumetric 3D stitching of optical coherence tomography volumes

Abstract Optical coherence tomography (OCT) is a noninvasive medical imaging modality, which provides highresolution transectional images of biological tissue. However, its potential is limited due to a relatively small field of view. To overcome this drawback, we describe a scheme for fully automated stitching of multiple 3D OCT volumes for panoramic imaging. The voxel displacements between two adjacent images are calculated by extending the Lucas-Kanade optical flow a lgorithm to dense volumetric images. A RANSAC robust estimator is used to obtain rigid transformations out of the resulting flow v ectors. T he i mages a re t ransformed into the same coordinate frame and overlapping areas are blended. The accuracy of the proposed stitching scheme is evaluated on two datasets of 7 and 4 OCT volumes, respectively. By placing the specimens on a high-accuracy motorized translational stage, ground truth transformations are available. This results in a mean translational error between two adjacent volumes of 16.6 ± 0.8 μm (2.8 ± 0.13 voxels). To the author’s knowledge, this is the first reported stitching of multiple 3D OCT volumes by using dense voxel information in the registration process. The achieved results are sufficient for providing high accuracy OCT panoramic images. Combined with a recently available high-speed 4D OCT, our method enables interactive stitching of hand-guided acquisitions.

Polynomial feature engineering for classification of textural images

I consider a number of methods of automatic quadratic features adjustment for digital textural images of biological tissues in order to improve the quality of classification. The proposed approaches are based on optimization procedures that use various quality criteria of a feature space as target functions. I investigate the methods based on random search, genetic algorithm, simulation of annealing, as well as the original hybrid algorithm. I presented results of experimental studies of the proposed algorithms on sets of real X-ray images of bone tissue and the lung CT images. We show that the hybrid algorithm provides more stable results regardless of the chosen quality criterion of the feature space, which is expressed in decreasing of the average percentage of incorrectly recognized images in comparison with the use of the specific optimization methods individually.

Coherent mid-infrared supercontinuum spectrum using a step-index tellurite fiber with all-normal dispersion

We demonstrate a coherent mid-infrared supercontinuum generation in a step-index tellurite fiber with an all-normal dispersion profile. A coherent supercontinuum spectrum spanning 1340–2840 nm in a 6.3-cm-long tellurite step-index fiber is obtained when the fiber is pumped with 200 fs laser pulses with a coupled peak power of 5.5 kW at 2000 nm. Such fiber-based coherent mid-infrared supercontinuum sources are highly required for the imaging of biological tissues in optical coherence tomography, pump–probe spectroscopy, pulse compression, and sensing.

Core-Shell Structures of Upconversion Nanocrystals Coated with Silica for Near Infrared Light Enabled Optical Imaging of Cancer Cells.

Optical imaging of cancer cells using near infrared (NIR) light is currently an active area of research, as this spectral region directly corresponds to the therapeutic window of biological tissues. Upconversion nanocrystals are photostable alternatives to conventional fluorophores. In our work, we have prepared upconversion nanocrystals of NaYF4:Yb/Er and encapsulated them in silica to form core-shell structures. The as-prepared core-shell nanostructures have been characterized for their structure, morphology, and optical properties using X-ray diffraction, transmission electron microscopy coupled with elemental mapping, and upconversion luminescence spectroscopy, respectively. The cytotoxicity examined using cell viability assay indicated a low level of toxicity of these core-shell nanostructures. Finally, these core-shell nanostructures have been utilized as photostable probes for NIR light enabled optical imaging of human breast cancer cells. This work paves the way for the development of advanced photostable, biocompatible, low-toxic core-shell nanostructures for potential optical imaging of biological cells and tissues.

Observation of short wavelength infrared (SWIR) Cherenkov emission.

Cherenkov emission induced by external beam radiation from a clinical linear accelerator has been shown in preclinical molecular imaging and clinical imaging. The broad spectrum Cherenkov emission should have a short wavelength infrared (SWIR, 1000-1700nm) component, as predicted theoretically. To the best of our knowledge, this Letter is the first experimental observation of this SWIR Cherenkov emission induced by external beam radiation. The measured spectrum of SWIR Cherenkov emission matches the theoretical prediction, with a fluence rate near one-third of the visible and near-infrared red emissions (Vis-NIR, 400-900nm). Imaging in water-based phantoms and biological tissues indicates that there is a sufficient fluence rate for radiotherapy dosimetry applications. The spatial resolution is improved approximately 5.3times with SWIR Cherenkov emission detection versus Vis-NIR Cherenkov emission, which provides some improvement in the potential for higher resolution Cherenkov emission dosimetry and molecular sensing during clinical radiotherapy by imaging with SWIR wavelengths.