Optic coherent tomography: a new high-resolution technology of visualization of tissue structures. Communication II. Optical images of benign and malignant entities

This is the second communication of a series of publications on Russian studies in the field of optical coherent tomography (OCT), the newest noninvasive highly resolving technology of visualization of the structure of biological tissues. By using the investing tissues as an example, this paper demonstrates the universal types of changes in their optical properties. Optimal images permit differentiate benign and malignant processes with a high degree of diagnostic accuracy. Diverse benign processes occurring in the epithelium are detected on the OCT images as changes in its height, the scattering properties and stroke of a basilar membrane. The absence of any structure on the image is the main OCT criterion for malignancy. The diagnostic efficiency of OCT is high in recognizing neoplasia of various mucous membranes: the sensitivity of the technique is 77-98%; its specificity and diagnostic accuracy are 71-96 and 81-87%, respectively.     

Full-field OCT

Optical coherence tomography (OCT) is an emerging technique for imaging of biological media with micrometer-scale resolution, whose most significant impact concerns ophthalmology. Since its introduction in the early 1990's, OCT has known a lot of improvements and sophistications. Full-field OCT is our original approach of OCT, based on white-light interference microscopy. Tomographic images are obtained by combination of interferometric images recorded in parallel by a detector array such as a CCD camera. Whereas conventional OCT produces B-mode (axially-oriented) images like ultrasound imaging, full-field OCT acquires tomographic images in the en face (transverse) orientation. Full-field OCT is an alternative method to conventional OCT to provide ultrahigh resolution images (approximately 1 microm), using a simple halogen lamp instead of a complex laser-based source. Various studies have been carried, demonstrating the performances of this technology for three-dimensional imaging of ex vivo specimens. Full-field OCT can be used for non-invasive histological studies without sample preparation. In vivo imaging is still difficult because of the object motions. A lot of efforts are currently devoted to overcome this limitation. Ultra-fast full-field OCT was recently demonstrated with unprecedented image acquisition speed, but the detection sensitivity has still to be improved. Other research directions include the increase of the imaging penetration depth in highly scattering biological tissues such as skin, and the exploitation of new contrasts such as optical birefringence to provide additional information on the tissue morphology and composition.     

Optical coherence tomography for ultrahigh resolution in vivo imaging

Optical coherence tomography (OCT) is an emerging biomedical optical imaging technique that performs high-resolution, cross-sectional tomographic imaging of microstructure in biological systems. OCT can achieve image resolutions of 1-15 microm, one to two orders of magnitude finer than standard ultrasound. The image penetration depth of OCT is determined by the optical scattering and is up to 2-3 mm in tissue. OCT functions as a type of 'optical biopsy' to provide cross-sectional images of tissue structure on the micron scale. It is a promising imaging technology because it can provide images of tissue in situ and in real time, without the need for excision and processing of specimens.     

Comparison of high frequency ultrasound and optical coherence tomography as modalities for high resolution and non invasive skin imaging.

High frequency ultrasound (HFUS) and optical coherence tomography (OCT) are techniques for high resolution imaging of tissues. The penetration depth of these modalities is limited, but it is sufficiently large enough for non invasive skin imaging. HFUS and OCT are based on the same concept. Waves (ultrasonic waves, respectively light waves) propagate along a narrow beam, are backscattered at tissue inhomogeneities and analyzed over time of flight to obtain spatially resolved morphological information. The objective of this paper is to compare HFUS and OCT in terms of resolution, dynamic range and contrast and to assess their value as tools for high resolution skin imaging. Measurements on phantoms and in vivo have been performed with a 100 MHz ultrasound system and an OCT-scanner working in the near infrared spectrum at 1300 nm wave-length. From the measurements, it can be concluded that OCT delivers an almost isotropic resolution (axial resolution about 5.8 microns, lateral resolution about 4.1 microns), whereas the resolution of the investigated HFUS system is more anisotropic (axial resolution about 9.3 microns, lateral resolution about 60 microns). HFUS and OCT show different penetration depths and a different contrast. Both techniques can, therefore, be combined advantageously in a multimodality approach to account for their individual characteristics.     

Optical coherence hyperspectral microscopy with a single supercontinuum light source

In the paper, we have developed an optical coherence hyperspectral microscopy with a single supercontinuum light source. The microscopy consists of optical coherence tomography (OCT) and hyperspectral imaging (HSI), which can visualize the structural and functional characteristics of biological tissues. The 500 to 700 nm band is selected for HSI and OCT imaging, where HSI enables imaging of oxygen saturation and hemoglobin (Hb) content, while OCT acquires structural characteristics to assess the morphology of biological tissues. The system performance of the optical coherence hyperspectral microscopy is verified by normal mice ears, and the practical applications of the microscopy is further performed in 4T1 and inflammation Balb/c mice ears in vivo. The experimental results demonstrate that the microscopy has potential to provide complementary information for clinical applications.     

Tissue proteomics using chemical immobilization and mass spectrometry

Proteomics analysis is important for characterizing tissues to gain biological and pathological insights, which could lead to the identification of disease-associated proteins for disease diagnostics or targeted therapy. However, tissues are commonly embedded in optimal cutting temperature medium (OCT) or are formalin-fixed and paraffin-embedded (FFPE) in order to maintain tissue morphology for histology evaluation. Although several tissue proteomic analyses have been performed on FFPE tissues using advanced mass spectrometry (MS) technologies, high-throughput proteomic analysis of OCT-embedded tissues has been difficult due to the interference of OCT in the MS analysis. In addition, molecules other than proteins present in tissues further complicate tissue proteomic analysis. Here, we report the development of a method using chemical immobilization of proteins for peptide extraction (CIPPE). In this method, proteins are chemically immobilized onto a solid support; interferences from tissues and OCT embedding are removed by extensive washing of proteins conjugated on the solid support. Peptides are then released from the solid phase by proteolysis, enabling MS analysis. This method was first validated by eliminating OCT interference from a standard protein, human serum albumin, where all of the unique peaks contributed by OCT contamination were eradicated. Finally, this method was applied for the proteomic analysis of frozen and OCT-embedded tissues using iTRAQ (isobaric tag for relative and absolute quantitation) labeling and two-dimensional liquid chromatography tandem mass spectrometry. The data showed reproducible extraction and quantitation of 10,284 proteins from 3996 protein groups and a minimal impact of OCT embedding on the analysis of the global proteome of the stored tissue samples.     

Optical coherence tomography for advanced screening in the primary care office

Optical coherence tomography (OCT) has long been used as a diagnostic tool in the field of ophthalmology. The ability to observe microstructural changes in the tissues of the eye has proved very effective in diagnosing ocular disease. However, this technology has yet to be introduced into the primary care office, where indications of disease are first encountered. We have developed a portable, handheld imaging probe for use in the primary care setting and evaluated its tissue site accessibility, ability to observe diseased tissue, and screening capabilities in in vivo human patients, particularly for pathologies related to the eye, ear and skin. Various stages of diabetic retinopathy were investigated using the handheld probe and early‐stage diabetic retinopathy was flagged as abnormal from the OCT images. At such early stages of disease, it is difficult to observe abnormalities with the limited tools that are currently available to primary care physicians. These results indicate that OCT shows promise to transform from being a diagnostic technology in the medical and surgical specialities to a screening technology in the primary care office and at the front‐line of healthcare. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Integrating photoacoustic microscopy with other imaging technologies for multimodal imaging

As a hybrid optical microscopic imaging technology, photoacoustic microscopy images the optical absorption contrasts and takes advantage of low acoustic scattering of biological tissues to achieve high-resolution anatomical and functional imaging. When combined with other imaging modalities, photoacoustic microscopy-based multimodal technologies can provide complementary contrast mechanisms to reveal complementary information of biological tissues. To achieve intrinsically and precisely registered images in a multimodal photoacoustic microscopy imaging system, either the ultrasonic transducer or the light source can be shared among the different imaging modalities. These technologies are the major focus of this minireview. It also covered the progress of the recently developed penta-modal photoacoustic microscopy imaging system featuring a novel dynamic focusing technique enabled by OCT contour scan.     

Liver tissue classification of en face images by fractal dimension‐based support vector machine

Full‐field optical coherence tomography (FF‐OCT) has been reported with its label‐free subcellular imaging performance. To realize quantitive cancer detection, the support vector machine model of classifying normal and cancerous human liver tissue is proposed with en face tomographic images. Twenty samples (10 normal and 10 cancerous) were operated from humans and composed of 285 en face tomographic images. Six histogram features and one proposed fractal dimension parameter that reveal the refractive index inhomogeneities of tissue were extracted and made up the training set. The other different 16 samples (8 normal and 8 cancerous) were imaged (190 images) and employed as the test set with the same features. First, a subcellular‐resolution tomographic image library for four histopathological areas in liver tissue was established. Second, the area under the receiver operating characteristics of 0.9378, 0.9858, 0.9391, 0.9517 for prediction of the cancerous hepatic cell, central vein, fibrosis, and portal vein were measured with the test set. The results indicate that the proposed classifier from FF‐OCT images shows promise as a label‐free assessment of quantified tumor detection, suggesting the fractal dimension‐based classifier could aid clinicians in detecting tumor boundaries for resection in surgery in the future.     

Affinity purification-mass spectrometry. Powerful tools for the characterization of protein complexes.

Multi-protein complexes are emerging as important entities of biological activity inside cells that serve to create functional diversity by contextual combination of gene products and, at the same time, organize the large number of different proteins into functional units. Many a time, when studying protein complexes rather than individual proteins, the biological insight gained has been fundamental, particularly in cases in which proteins with no previous functional annotation could be placed into a functional context derived from their 'molecular environment'. In this minireview, we summarize the current state of the art for the retrieval of multiprotein complexes by affinity purification and their analysis by mass spectrometry. The advances in technology made over the past few years now enable the study of protein complexes on a proteomic scale and it can be anticipated that the knowledge gathered from such projects will fuel drug target discovery and validation pipelines and that the technology is also going to prove valuable in the emerging field of systems biology.     

Noninvasive cross-sectional observation of three-dimensional cell sheet-tissue-fabrication by optical coherence tomography

Cell sheet engineering allows investigators/clinicians to prepare cell-dense three-dimensional (3-D) tissues, and various clinical trials with these fabricated tissues have already been performed for regenerating damaged tissues. Cell sheets are easily manipulated and 3-D tissues can be rapidly fabricated by layering the cell sheets. This study used optical coherence tomography (OCT) to noninvasively analyze the following processes: (1) adhesions between layered cell sheets, and (2) the beating and functional interaction of cardiac cell sheet-tissues for fabricating functional thicker 3-D tissues. The tight adhesions and functional couplings between layered cell sheets could be observed cross-sectionally and in real time. Importantly, the noninvasive and cross-sectional analyses of OCT make possible to fabricate 3-D tissues by confirming the adherence and functional couplings between layered cell sheets. OCT technology would contribute to cell sheet engineering and regenerative medicine.     

微针及其在生物诊疗中的应用研究进展

微针在过去几十年中发展迅速,在透皮给药等领域已得到了广泛的研究.近些年随着微电子技术的蓬勃兴起,微针在生物诊疗中的应用越来越多的受到人们关注.通过微针提取血液和组织液进行检测分析以及微针作为电极在体液内直接对血糖、黑色素瘤、pH值等进行实时检测方面,都展现出了良好的实时检测应用前景.文中综述了微针的材料与结构设计及其在...     

Review of russian literature on biological action of DC and low-frequency AC magnetic fields

This review considers the Russian scientific literature on the influence of weak static and of low-frequency alternating magnetic fields on biological systems. The review covers the most interesting works and the main lines of investigation during the period 1900 to the present. Shown here are the historical roots, beginning with the ideas of V. Vernadsky and A. Chizhevsky, which led in the field of Russian biology to an increasing interest in magnetic fields, based on an intimate connection between solar activity and life on the Earth, and which determined the peculiar development of Russian magnetobiology. The variety of studies on the effects of magnetic storms and extremely low-frequency, periodic variations of the geomagnetic field on human beings and animals as well as on social phenomena are described. The diverse experiments involving artificial laboratory magnetic fields acting on different biological entities under different conditions are also considered. A series of theoretical advances are reviewed that have paved the way for a step-by-step understanding of the mechanisms of magnetic field effects on biological systems. The predominantly unfavorable influence of magnetic fields on living beings is shown, but the cases of favorable influence of magnetic fields on human beings and lower animals are demonstrated as well. The majority of Russian investigations in this area of science has been unknown among the non-Russian speaking audience for many reasons, primarily because of a language barrier. Therefore, it is hoped that this review may be of interest to the international scientific community.     

Spectrally encoded coherence tomography and reflectometry: Simultaneous en face and cross‐sectional imaging at 2 gigapixels per second

Non‐invasive biological imaging is crucial for understanding in vivo structure and function. Optical coherence tomography (OCT) and reflectance confocal microscopy are two of the most widely used optical modalities for exogenous contrast‐free, high‐resolution, three‐dimensional imaging in non‐fluorescent scattering tissues. However, sample motion remains a critical barrier to raster‐scanned acquisition and reconstruction of wide‐field anatomically accurate volumetric datasets. We introduce spectrally encoded coherence tomography and reflectometry (SECTR), a high‐speed, multimodality system for simultaneous OCT and spectrally encoded reflectance (SER) imaging. SECTR utilizes a robust system design consisting of shared optical relays, scanning mirrors, swept laser and digitizer to achieve the fastest reported in vivo multimodal imaging rate of 2 gigapixels per second. Our optical design and acquisition scheme enable spatiotemporally co‐registered acquisition of OCT cross‐sections simultaneously with en face SER images for multivolumetric mosaicking. Complementary axial and lateral translation and rotation are extracted from OCT and SER data, respectively, for full volumetric estimation of sample motion with micron spatial and millisecond temporal resolution.      

Fertility factors in Pseudomonas putida: selection and properties of high-frequency transfer and chromosome donors.

The octane plasmid (OCT) in Pseudomonas putida strains has been shown to be transferred at low frequency. However, bacteria which had newly received this plasmid showed a transient increase in donor ability. Using Octane+ P. putida as the donor, the transfer of most chromosomal markers was shown to be independent of OCT transfer, whereas the mobilization of the octanoate catabolism genes (octanoic and acetate) was dependent on OCT plasmid transfer. The presence of a fertility factor termed FPo has been postulated to explain these results. Strains carrying only this fertility factor have been obtained from strains carrying both OCT and FPo plasmids. Strains in which the OCT plasmid was transferred at high frequencies have also been isolated, and chromosome mobilization by OCT and FPo has been compared. A different gradient of transmission by OCT and FPo has been observed. It has also been shown that chromosome transfer by OCT was dependent on the bacterial recombination system, whereas the chromosome transfer by FPo was unaffected by the presence of a rec mutation in the donor strain.     

OCT于大鼠脱水组织中光散射特性的分析研究

目的:探讨活体组织液体含量的改变对OCT成像的影响,以期提高OCT在诊断组织病理性质方面的能力。方法:实验中复制脱水大鼠病理模型,应用光学相干层析成像设备,进行大鼠舌浅表组织在体显微成像检测,并对图像中组织的信号衰减特性进行量化分析。结果:正常对照组大鼠体重明显增加,病理模型组显著下降,病理模型组于脱水3天和5天后组织的平均OCT信号衰减系数明显高于正常对照组(P<0.01),且5天较3天的病理模型组组织的信号衰减系数变化尤其显著(P<0.01)。结论:改变组织含液量,可显著改变OCT成像效果,且通过对OCT图像中信号的衰减系数分析,可获得组织细微的散射变化,从而有望提高OCT技术在组织性质方面的诊断能力。     

Study of the Morphological and Functional State of Higher Plant Tissues by Optical Coherence Microscopy and Optical Coherence Tomography

Comparative analysis of two optical methods—optical coherence tomography (OCT) and optical coherence microscopy (OCM)—was made for vital visualization of plant tissues in tomato (Lycopersicon esculentum Mill), spiderwort (Tradescantia pallida (Rose) D. Hunt), orach (Atriplex sp.), and leaves and seeds of medium starwort (Stellaria media L.). The obtained OCT- and OCM-images allowed the morphological and functional state of plant tissues to be assessed in vivo. A higher spatial resolution of the OCM method, as compared to OCT method, allowed plant morphological structures to be identified with greater confidence. The morphological and functional state of tissues can be monitored with a time resolution of 1–4 s in intact plants, without removing them from the habitat.__________Translated from Fiziologiya Rastenii, Vol. 52, No. 4, 2005, pp. 628–634.Original Russian Text Copyright © 2005 by Kutis, Sapozhnikova, Kuranov, Kamenskii.     

High-resolution imaging of proteins in human teeth by scanning probe microscopy

High-resolution studies of dental tissues are of considerable interest for biomedical engineering and clinical applications. In this paper, we demonstrate the application of piezoresponse force microscopy (PFM) to nanoscale imaging of internal structure of human teeth by monitoring the local mechanical response to an electrical bias applied via a conductive tip. It is shown that PFM is capable of detecting dissimilar components of dental tissues, namely, proteins and calcified matrix, which have resembling morphology but different piezoelectric properties. It is demonstrated that collagen fibrils revealed in chemically treated intertubular dentin exhibit high piezoelectric activity and can be visualized in PFM with spatial resolution of 10 nm. Evidence of the presence of protein inclusions of 100-200 nm wide and several micrometers long in tooth enamel has been obtained. Furthermore, it is found that the peritubular dentin and intertubular dentin exhibit different piezoelectric behavior suggesting different concentration of collagen fibrils. The obtained results demonstrate a high potential of PFM in providing an additional insight into the structure of dental tissues. It is suggested that the PFM approach can be used to study the structure of a wide range of biological materials by monitoring their electromechanical behavior at the nanoscale.     

Dependence of optical attenuation coefficient and mechanical tension of irradiated human cartilage measured by optical coherence tomography

As banked human tissues are not widely available, the development of new non-destructive and contactless techniques to evaluate the quality of allografts before distribution for transplantation is very important. Also, tissues will be processed accordingly to standard procedures and to minimize disease transmission most tissue banks will include a decontamination or sterilization step such as ionizing radiation. In this work, we present a new method to evaluate the internal structure of frozen or glycerol processed human cartilages, submitted to various dosis of irradiation, using the total optical attenuation coefficient retrieved from optical coherence tomography (OCT) images. Our results show a close relationship between tensile properties and the total optical attenuation coefficient of cartilages. Therefore, OCT associated with the total optical attenuation coefficient open a new window to evaluate quantitatively biological changes in processed tissues.