New insights into plant cell walls by vibrational microspectroscopy

Vibrational spectroscopy provides non-destructively the molecular fingerprint of plant cells in the native state. In combination with microscopy, the chemical composition can be followed in context with the microstructure, and due to the non-destructive application, in-situ studies of changes during, e.g., degradation or mechanical load are possible. The two complementary vibrational microspectroscopic approaches, Fourier-Transform Infrared (FT-IR) Microspectroscopy and Confocal Raman spectroscopy, are based on different physical principles and the resulting different drawbacks and advantages in plant applications are reviewed. Examples for FT-IR and Raman microscopy applications on plant cell walls, including imaging as well as in-situ studies, are shown to have high potential to get a deeper understanding of structure–function relationships as well as biological processes and technical treatments. Both probe numerous different molecular vibrations of all components at once and thus result in spectra with many overlapping bands, a challenge for assignment and interpretation. With the help of multivariate unmixing methods (e.g., vertex components analysis), the most pure components can be revealed and their distribution mapped, even tiny layers and structures (250 nm). Instrumental as well as data analysis progresses make both microspectroscopic methods more and more promising tools in plant cell wall research.     

The Challenge of Spectroscopy in the Microanalysis of Biological Surfaces

Abstract

Chemical interactions between solids and their environments are initially characterized by the chemical composition of the surfaces. This is especially true for biochemical processes, such as ionic transport through biological membranes, the maintenance of mineral ion homeostasis which is critically dependent on the activity of the “bone membrane,” the biocompatibility of surgical implants which appears to be only a surface problem, etc. This is because biological surfaces have shown an increased interest for different fields of science. However, biological surfaces are heterogeneous and generally more complex than inorganic ones, and biological subsurfaces (internal surfaces) are not usually identical with the external surfaces. Taking into account that much of the functional significance of biological surfaces is related to the internal surfaces and these are normally hidden, we must expose them for analysis without introducing artifacts during the preparative procedures. As a result, the specimen preparation techniques are to a large extent the first limiting factor. Preparation of fully mineralized tissues such as teeth, bone, and kidney stones consisting of embedding in resin, sectioning, and fine polishing have been successfully accomplished. However, soft biological materials are prepared by cryogenic methods to preserve the original elemental distributions (but altering the tissue morphology) or by chemical fixation and water replacement by embedding media to preserve the morphology (but increasing diffusion and translocation of elements).     

山东肿足蕨的红外光谱分析

本文首次利用傅里叶红外光谱法对山东肿足蕨不同部位和全草的红外光谱进行了对比分析。分析表明, 同株山东肿足蕨不同部位的红外光谱中, 它们吸收峰的峰形、峰位大体相同, 但由于它们化学成分和相对含量出现差异, 红外光谱的峰形、峰位和峰相对强度存在差异。文中还根据各种化学键和官能团振动所引起的吸收峰形状和范围, 判断推理出山东肿足蕨中所含的化学成分。山东肿足蕨的红外光谱分析, 为道地药材山东肿足蕨的开发应用提供了科学依据。     

Effects of soft X‐ray radiation damage on paraffin‐embedded rat tissues supported on ultralene: a chemical perspective

Radiation damage is an important aspect to be considered when analysing biological samples with X‐ray techniques as it can induce chemical and structural changes in the specimens. This work aims to provide new insights into the soft X‐ray induced radiation damage of the complete sample, including not only the biological tissue itself but also the substrate and embedding medium, and the tissue fixation procedure. Sample preparation and handling involves an unavoidable interaction with the sample matrix and could play an important role in the radiation‐damage mechanism. To understand the influence of sample preparation and handling on radiation damage, the effects of soft X‐ray exposure at different doses on ultralene, paraffin and on paraffin‐embedded rat tissues were studied using Fourier‐transform infrared (FTIR) microspectroscopy and X‐ray microscopy. Tissues were preserved with three different commonly used fixatives: formalin, glutaraldehyde and Karnovsky. FTIR results showed that ultralene and paraffin undergo a dose‐dependent degradation of their vibrational profiles, consistent with radiation‐induced oxidative damage. In addition, formalin fixative has been shown to improve the preservation of the secondary structure of proteins in tissues compared with both glutaraldehyde and Karnovsky fixation. However, conclusive considerations cannot be drawn on the optimal fixation protocol because of the interference introduced by both substrate and embedding medium in the spectral regions specific to tissue lipids, nucleic acids and carbohydrates. Notably, despite the detected alterations affecting the chemical architecture of the sample as a whole, composed of tissue, substrate and embedding medium, the structural morphology of the tissues at the micrometre scale is essentially preserved even at the highest exposure dose.     

Arsenic-Induced Biochemical Changes in Labeo rohita Kidney: An FTIR Study

ABSTRACT

Arsenic is a toxic heavy metal that occurs naturally in water, soil, and air. It is widespread in the environment as a consequence of both anthropogenic and natural processes. In the current study, an attempt has been made to analyze the arsenic-induced molecular changes in macromolecular components like proteins and lipids in the kidney tissues of edible fish Labeo rohita using Fourier transform infrared (FTIR) spectroscopy. The FTIR spectrum of kidney tissue is quite complex and contains several bands arising from the contribution of different functional groups. The detailed spectral analyses were performed in three distinct wave number regions, namely 3600–3050 cm^?1, 3050–2800 cm^?1, and 1800–800 cm^?1. The current study shows that the kidney tissues are more vulnerable to arsenic intoxication. FTIR spectra reveal significant differences in both absorbance intensities and areas between control and arsenic-intoxicated kidney tissues; this result indicates that arsenic intoxication induces significant alteration on the major biochemical constituents such as lipids and proteins and leads to compositional and structural changes in kidney tissues at the molecular level. The current study confirms that FTIR spectroscopy can be successfully applied to toxicologic and biological studies.     

Applications of spectral analysis in medical ultrasonography.

Spectral analysis of ultrasonic reflections from biological tissues can be used to determine basic tissue parameters for use in differential diagnosis. This paper describes the use of the technique under circumstances encountered in several types of clinical examinations. The applications are illustrated with results obtained from laboratory measurements with a system now being employed in a clinical evaluation programme. The test objects studied simulate tissues with planar boundaries, tissues with heterogeneous interior structure, and tissues causing acoustic 'shadowing' of posterior regions.     

Polarimetry group theory analysis in biological tissue phantoms by Mueller coherency matrix

The characterization of biological tissues by optical techniques provides several advantages over other techniques. Optical techniques enable to perform high resolution and contrast imaging, in a non-invasive way and with no-contact. Biological tissues are turbid media that strongly scatter light. The ultrastructure of some tissues makes them present a certain degree of anisotropy. Both scattering and anisotropy affect light polarization. Some pathologies alter these characteristics of the tissue. As a consequence polarized light can be used to extract additional information and achieve a better diagnosis.In this work, Group Theory is applied to analyse the polarization behavior of several samples. Firstly, the Mueller matrix for each sample is measured. Then, the Mueller Coherency matrix is obtained by means of the SU(4)-O + (6) homomorphism. Finally, the target decomposition theorem is applied by analyzing the eigenvalues and eigenvectors, and subsequently the different polarimetric effects are separated. In this way, the contrast of tissue imaging can be increased. This analysis is applied to biological tissue phantoms, which consisted on glucose suspensions of polystyrene spheres with different scatterer concentrations. Their behaviour can be modeled by means of single or multiple scattering depending on the concentration, either in the Rayleigh or Mie regimes. The same procedure could be used in a wide range of applications, like the study of cancerous cells that grow without control in cell cultures, or erythrocytes monitoring in anemia. The technique also has a great potential to be applied in Polarization Sensitive Optical Coherence Tomography (PS-OCT).     

Estimation of optical abnormalities in breast phantom by diffuse equation

Optical imaging is an emerging method for bio-imaging. The advantages of this imaging provide non-ionizing and safe radiation, non-invasive and functional medical imaging. Due to diffusion of photons inside biological tissues, its image processing is complicated. So in spite of these advantages, this imaging method has not been progressed like ultrasound imaging and magnetic resonance imaging (MRI). Also, the penetration depth of photons inside biological phantom is low. To overcome this problem, the complicated diffusion of photons through tissue must be modeled. The diffuse equation can be applied to simulate photons through turbid media like biological tissues. In this paper, the diffuse equation is used to study propagation of diffuse photons. The green function method is applied to solve this equation, and then the optical properties of abnormalities in breast phantom are estimated. This fast method can be applied for image processing.     

Carotenes and carotenoids in natural biological samples: a Raman spectroscopic analysis

Raman spectroscopic studies of a range of naturally occurring carotenoids in over 50 specimens of plant tissue and a range of standard extracts have been undertaken, and the characteristic bands of CC and C C stretching and C CH bending have been recorded. Comparison of the spectroscopic data with the chemical assignment of the carotenoids from chemical extraction of the plant tissues reveals that there is a problem in the interpretation of the spectroscopic data which can be attributed to significant wavenumber shifts, particularly in the CC stretching band wavenumber, for carotenoids in the organic tissues arising from molecular interactions between the carotenoid and its host matrix. The simple identification of carotenoids in biological tissues on the basis of comparison with the standard spectra of extracted material must be made with caution; the progressive shift in wavenumber of the CC stretching band in the conjugated polyene chain of carotenoids with the number of CC groups, and hence the identification of the carotenoid, cannot be unambiguously interpreted for the range of materials studied here. Copyright © 2009 John Wiley & Sons, Ltd.     

时域光声谱技术及其在生物组织检测中的应用

当强度调制的光束照射于吸收物质,周期性热流使周围的介质热胀冷缩而激发声波,这种将光能转化为声能的现象称为光声效应。基于光声效应的时域光声谱技术将光学和声学有机地结合,为生物组织的无损检测技术提供了新的检测手段。该技术能够实现类似光学技术的高对比度和近似于声学技术的高精度和穿透深度,在生物医学检测中具有广阔的应用前景。文章介绍时域光声谱技术的原理及其在生物组织成分检测和层析成像检测中的应用。     

乳腺组织形态基元共焦显微拉曼光谱的研究

利用共焦显微拉曼方法,测定了乳腺组织切片的Mapping拉曼谱,光谱采样体积达到μm尺度.通过对这些共焦拉曼谱与商业提纯肌动蛋白、胶原等生化物质做出的谱的分析比较,分离出了来自细胞质、细胞间质、脂肪以及乳腺沉积物这些组织形态基元的拉曼谱;用K阶簇分析方法,还获得了细胞核基元谱;基元谱与提纯化学物质谱的相关分析印证了组织形态基元谱的来源.还分析总结了这些组织形态基元谱的特征.这些工作为深入理解组织形态与其拉曼谱的关系以及乳腺组织形态拉曼模型的建立奠定了基础.     

Investigation of oscillations of biological tissues and their phantoms with model defects

The potential to create a remote, contactless, diagnostic method for biological tissues or materials with properties similar to those of biological tissues, so-called phantoms, is studied. The method is based on measurement of local oscillations of the surface of material under study under the action of acoustic and ultrasonic waves. The application of a laser-scanning vibrometer provides a means for visualization oscillations of a biological tissue phantom with model defects in the form of air bubbles, regions with higher density, and laminations, and allows determination of the positions of these defects.     

Atomic force microscopy for the investigation of molecular and cellular behavior

The present review details the methods used for the measurement of cells and their exudates using atomic force microscopy (AFM) and outlines the general conclusions drawn by the mechanical characterization of biological materials through this method. AFM is a material characterization technique that can be operated in liquid conditions, allowing its use for the investigation of the mechanical properties of biological materials in their native environments. AFM has been used for the mechanical investigation of proteins, nucleic acids, biofilms, secretions, membrane bilayers, tissues and bacterial or eukaryotic cells; however, comparison between studies is difficult due to variances between tip sizes and morphologies, sample fixation and immobilization strategies, conditions of measurement and the mechanical parameters used for the quantification of biomaterial response. Although standard protocols for the AFM investigation of biological materials are limited and minor differences in measurement conditions may create large discrepancies, the method is nonetheless highly effective for comparatively evaluating the mechanical integrity of biomaterials and can be used for the real-time acquisition of elasticity data following the introduction of a chemical or mechanical stimulus. While it is currently of limited diagnostic value, the technique is also useful for basic research in cancer biology and the characterization of disease progression and wound healing processes.     

Retrieval of chromophore concentration in a tissue phantom by diffuse reflectance spectroscopy

This study describes a method for analysis of back reflectance spectroscopic data to estimate the concentration of endogenous or exogenous chromophores of tissue, noninvasively and in real time. In the study, tissue phantoms were prepared using intralipid and two chromophores, indocyanine green and methylene blue. Reduced scattering and absorption coefficients ranges of tissue phantoms were kept in the range of biological tissues. Spectroscopic measurements on the tissue phantoms were carried out using a miniature spectrometer, an optical fiber probe, a halogen tungsten light source and a laptop. Monte Carlo simulations of the experiments were run, and an average optical path lengths of the detected photons were obtained for different absorption and scattering coefficients. The average optical path lengths of the photons were used to estimate concentrations of the chromophores in the tissue phantoms. Scattering and absorption coefficients were estimated with an average error of 4.7 and 5.4%, respectively. The developed method has the potential to be used in diagnosis of pathologic tissues based on the variation of biochemical composition of tissues and in photodynamic therapy to estimate the concentration of photosensitizers.     

Fourier transform infrared spectroscopy (FTIR) application chemical characterization of enamel,dentin and bone

Fourier transform infrared spectroscopy (FTIR) has been used extensively for chemical characterization of mineralized tissues in the past few decades. FTIR is an ideal technique to analyze chemical structural properties of natural materials, since the frequencies of several vibrational modes of organic and inorganic molecules are active in the infrared. This review discusses the use of FTIR methodology, highlighting the attenuated total reflection (ATR) sampling mode, particularly for characterization of enamel, dentin and bone tissues. Enamel, dentin and bone, are composed of an organic and a mineral phase. The mineral phase is characterized essentially as nonstoichiometric substituted apatite, being the carbonate and phosphate spectral peaks the main representative of these phase. Organic matrix of the post-eruptive enamel is small (～1% weight (wt)). The dentin and bone organic phases are mainly composed of type I collagen that appears as spectral bands of amide I, amide II, amide III bands. Furthermore, synthetic apatite materials are being designed for total or partial replacement, restoration or augmentation of these biological tissues with FTIR assistance.     

基于生物阻抗谱成像的生物组织检测方法

基于电阻抗成像技术和生物阻抗谱技术,提出一种面向生物组织检测的生物阻抗谱成像方法.该方法将目标区域可视化并精准识别目标种类,可用于肺癌早期检测,协助临床医生对早期肺癌进行精准检测,提高早期肺癌的治愈率.本文通过数值仿真的方法验证生物阻抗谱成像方法在肺癌早期检测中的可行性和有效性,仿真结果表明:1)生物阻抗谱成像方法可以实现早期肺癌区域的可视化,并精确判别出早期肺癌种类; 2)生物阻抗谱成像方法中阻抗谱的最佳采集模式是4次循环采集,最佳分类器是Linear-SVM, 5折交叉验证的平均分类准确率可以达到99.9%.为了验证仿真结果,本文选取3种具有不同电学特性的生物组织模拟癌变区域进行了检测.实验结果表明该方法可以对生物组织区域可视化,并判别出生物组织的种类.该方法可以兼顾电阻抗成像和生物阻抗谱方法的优点,有望用于肺癌早期检测.     

Interaction of biological and nonbiological surfaces. New opportunities for research and technologies using synchrotron radiation

The applicability of synchrotron radiation to implementation of medical ideas associated with introduction of nonbiological objects into a human body—implants, drug nanocapsules, and X-ray therapeutic means (metal nanoparticles and nanocrystalline phosphors and scintillators)—is considered. Synchrotron radiation presents new possibilities of analyzing the surface with a resolution comparable to the sizes of biological nanostructures involved in interactions such as chemical and biochemical modification of implant surfaces in vitro; activation of implant surface integration with biological tissue in a tissue culture (in vitro) and in vivo; biochemical modification of therapeutic nanoparticle surface in vitro; immunocamouflage by host proteins; attachment of “molecular targets”, i.e., antibodies, to target tissue to provide targeted delivery vehicles; and local activation of X-ray therapeutic drugs and drug nanocapsules in biological tissues. A functional block diagram of a medical technological station is given.     

Comparing Diffusion Approximation with Radiation Transfer Analysis for Light Transport in Tissues

The diffusion model that is an approximation of the equation of radiation transfer is typically used to describe photon migration in scattering-dominant media. In general biological tissue is highly scattering and very weakly absorbing against near-infrared light, yet it is heterogeneous and may contain relatively highly absorbing or low-scattering regions. Here applicability of the diffusion approximation over the radiative transfer theory for describing ultrafast laser transport in biological tissues is numerically studied and investigated over different kinds of tissue conditions and geometries. Tissues having tumors of different sizes, locations and nature as well as dual-tumor and low-scattering conditions are considered. Radiation transfer analysis is taken as a comparison objective and it is initially proved to be accurate in benchmark comparisons with Monte Carlo simulation. The results predict systematically about the compatible conditions where and when we can use the diffusion approximation and the conditions in which the diffusion approximation may provide misleading results.     

Ultrasound elastomicroscopy using water jet and osmosis loading: potentials for assessment for articular cartilage

Research in elasticity imaging typically relies on 1-10 MHz ultrasound. Elasticity imaging at these frequencies can provide strain maps with a resolution in the order of millimeters, but this is not sufficient for applications to skin, articular cartilage, or other fine structures. In this paper, we introduced two methods of ultrasound elastomicroscopy using water jet and osmosis loading for imaging the elasticity of biological soft tissues with high resolutions. In the first system, the specimens were compressed using water jet compression. A water jet was used to couple a focused 20 MHz ultrasound beam into the specimen and meanwhile served as a "soft" indenter. Because there was no additional attenuation when propagating from the ultrasound transducer to the specimen, the ultrasound signal with high signal-to-noise ratio could be collected from the specimens simultaneously with compressing process. The compression was achieved by adjusting the water flow. The pressure measured inside the water pipe and that on the specimen surface was calibrated. This system was easily to apply C-scan over sample surfaces. Experiments on the phantoms showed that this water jet indentation method was reliable to map the tissue stiffness distribution. Results of 1D and 2D scanning on phantoms with different stiffness are reported. In the second system, we used osmotic pressure caused by the ion concentration change in the bathing solutions for the articular cartilage to deform them. When bovine articular cartilage specimens were immerged in solutions with different salt concentration, a 50 MHz focused ultrasound beam was used to monitor the dynamic swelling or shrinkage process. Results showed that the system could reliably map the strain distribution induced by the osmotic loading. We extract intrinsic layered material parameters of the articular cartilage using a triphasic model. In addition to biological tissues, these systems have potential applications for the assessment of bioengineered tissues, biomaterials with fine structures, or some engineering materials. Further studies are necessary to fully realize the potentials of these two new methods.     

Source of reaction-diffusion coupling in confined systems due to temperature inhomogeneities

Diffusion is often accompanied by a reaction or sorption which can induce temperature inhomogeneities. Monte Carlo simulations of Lennard-Jones atoms in zeolite NaCaA are reported with a hot zone presumed to be created by a reaction. Our simulations show that localized hot regions can alter both the kinetic and transport properties. Further, enhancement of the diffusion constant is greater for larger barrier height, a surprising result of considerable significance to many chemical and biological processes. We find an unanticipated coupling between reaction and diffusion due to the presence of hot zone in addition to that which normally exists via concentration.