生物组织折射率的概念与测量方法评述

折射率作为描述介质光学特性的一个重要物理参量,它在组织光学理论中起着非常重要的作用.随着组织光学研究的深入,生物组织折射率的概念也在不断地完善和发展.本文将主要介绍生物组织折射率的基本概念和测量手段的研究进展情况,明确了生物组织折射率的概念,评述了现有测量方法的优缺点,希望为生物组织折射率问题的进一步研究提供有益参考.     

反卷积在生物组织光传输特性研究中的应用

生物组织中的光传输特性可以以点扩散函数表征,即在线状光束入射条件下,生物组织中某一深度层面上的光强度场分布。为获得点扩散函数的具体形式,已发展了多种理论分析方法,其中以Monte Carlo模拟方法最具代表性。但现有理论计算方法都要以生物组织的光学参数已知为前提,而光学参数的准确度直接影响着计算的精度。从线性平移不变系统理论出发,生物组织内一定深度层面上的光强分布被看成是光源强度分布与点扩散函数的卷积,从而提出通过测量在轴对称的准直扩展光源照射条件下,组织中特定层面上的光强度分布,利用反卷积重建生物组织的点扩散函数的方法,并将这种方法应用于典型生物组织透射面上点扩散函数的重建,得到了相应的点扩散函数。实验结果与Monte Carlo模拟的结果吻合较好,表明该方法从实验上获得生物组织点扩散函数的正确性和有效性。     

传递函数在生物组织光传输问题中的应用

本文将传递函数的概念引入生物组织光传输问题,并将传递函数理论用于面光源照射下生物组织内特定深度层面上光场强度分布的理论计算。结合Monte Carlo模拟获取脉冲相应函数,我们分析了不同面光源照射下层状组织样品透射面上的光场强度分布。理论计算结果与实验测试结果的一致性较好,这充分说明了本文建立的基于Monte Carlo模拟的传递函数方法是一种处理面光源照射下生物组织内光场空间的直接而有效的手段。     

一种微距测量组织光学参数的方法

测量生物组织的光学参数是近红外光学检测(NIR)的主要任务之一,然而常规的近红外光学检测由于受扩散近似的限制,测量的距离一般大于2cm-3cm。本文从提高近红外光学检测空间分辨率的角度,设计了一种可用于测量生物组织局部、浅表光学参数的方法。通过使用光纤探针,实现了在微小光源-探测器间距时(＜1mm)测量组织光学参数。模型实验的结果表明该方法可反映生物组织中吸收系数及散射系数的变化率。     

近红外无创伤血糖测量的组织光学基础研究

人体血糖浓度无创伤测量是当今学术界和医学界普遍关注的课题。在分析了血糖浓度无创伤测量的意义、现有的测量方法及其进展后,从组织光学角度分析血糖浓度无创伤测量中包含的研究内容,并根据当前的研究现状,提出血糖浓度无创伤测量所存在的问题。通过组织光学角度对血糖浓度无创伤测量方法的剖析,更加明确血糖测量的研究任务,有望促使其更进一步的发展以及血液中其他成分的无创测量。     

采用OCT系统对组织光学通透的研究

组织通透方法采用高折射率化学试剂对生物组织进行渗透,改变组织的光学均匀性,可以有效地改善光学成像的穿透深度,受到生物医学光学研究领域的重视。利用光学相干层析成像技术,测量通透过程中不同测量深度下组织的散射特征的变化。通过采用系统信号对数的梯度值近似地表征光学散射系数,研究了通透过程中组织的散射特征随渗透时间和测量深度的动态关系。实验证明了组织通透可以有效地增加光子的穿透深度,并改善成像质量。研究发现:不同测量深度处组织的散射系数及其变化幅度、变化过程和变化趋势等均存在一定的差异性,并与组织的微观结构、其通透效果,化学试剂在组织中的渗透行为等有密切关系,有助于组织通透过程的理解,并为组织通透机制提供可能的实验依据。     

激光照射血液疗法中的组织光学研究及其应用

本文从组织光学的角度 ,对激光照射血液疗法中的发展现状与存在问题进行分析 ,提出该领域应开展的若干研究内容。本文开展了激光照射血液疗法所涉及的若干组织光学研究。获得了中国人血液的吸收系数、散射系数、各项异性因子、全衰减系数等 ;研究获得了人血管的若干光学参数。通ＭｏｎｔｅＣａｒｌｏ模拟计算 ,以及实验测试结果 ,分别从理论与实际上分析对比了激光血管内照射时 ,光在人体血管、血液中的分布情况 ,以及不同的激光入射角 ,发散角 ,不同的血管直径等因素对激光血液照射的影响。根据对应的组织光学的研究结果 ,文中还讨论并提出…     

用于Monte Carlo模拟的复杂生物组织模型

Monte Carlo（MC）模拟被广泛应用于光子在生物组织中的传输研究。通常模拟时将生物组织近似为均匀的平板分层介质,当层状生物组织中含有异常物质（如肿瘤细胞等）或正常生物组织为非平板的复杂结构时,其模拟中的组织模型将会有相应的改变。通过探讨这几类生物组织的MC模拟模型,总结并分析模型建立的关键问题,对基于MC模拟的各种生物组织光学检测研究提供了指导。     

生物组织光学特性的时间分辨透射测量

基于时域漫射理论,提出一种用透射光来确定生物组织光学特性的方法。通过对生物组织的模拟测量,结果表明：此方法具有算法简单,速度快,精度高等优点,并比较了透射和反射两种测量法的精度。     

生物组织的超声调制光学成像技术

具有超声定位的高空间分辨率和光学检测的高灵敏度的超声调制光学成像技术是一种有前途的无损的生物组织成像技术。文章阐述了该技术的成像原理,评述了前人在散射介质中声光作用机制的理论研究;介绍了该领域在技术路线上的最新研究进展;最后总结了超声调制光学成像技术的优点并展望了其在生物医学领域的应用前景。     

生物组织的折射和折射率

光在生物组织中的传播与组织的光学性质有关。光通过组织时,光强和光的偏振状态会发生变化。而折射率是组织光学用来评价组织改变光线行进方向的基本参量。本文以菲涅耳公式为理论依据,用空气一组织界面的反射率、生物组织薄膜的反射率和生物组织反射光的倔振分量,推算生物组织的折射率。     

Optical clearing for multiscale biological tissues

Three‐dimensional reconstruction of tissue structures is essential for biomedical research. The development of light microscopes and various fluorescent labeling techniques provides powerful tools for this motivation. However, optical imaging depth suffers from strong light scattering due to inherent heterogeneity of biological tissues. Tissue optical clearing technology provides a distinct solution and permits us to image large volumes with high resolution. Until now, various clearing methods have been developed. In this study, from the perspective of the end users, we review in vitro tissue optical clearing techniques based on the sample features in terms of size and age, enumerate the methods suitable for immunostaining and lipophilic dyes and summarize the combinations with various imaging techniques. We hope this review will be helpful for researchers to choose the most suitable clearing method from a variety of protocols to meet their specific needs.      

Spectral attenuation of brain and retina tissues in the near‐infrared range measured using a fiber‐based supercontinuum device

A novel setup for the efficient constant optical measurements of biological tissues in the near infrared is presented. The system combines the use of a fiber‐based supercontinuum source with a simple optics fiber collimator. This configuration allows a wide spectral range of measurement and, at the same time, can efficiently filter the straightforward transmitted light while avoiding scattered light. As a performance example, the optical characterization of rat brain and retina tissues are shown. The attenuation coefficient for both tissues in the near infrared region is also obtained. This technique could be applied in clinical research as a noninvasive method with several potential practical applications.     

Sampling,storage, and preparation of biological material for trace-element analysis

Sampling, storage, and preparation of biological material for analysis are some of the most important factors in the study of trace elements in biomedical fields. If due care is not given to these factors, the resulting data may often be meaningless, regardless of the analytical technique chosen for analysis. A survey of the data in the literature would support this view, in which wide differences in the trace element composition of biological tissues have been observed, and not all these differences are attributed to variations in the biological systems. In this paper, the following points are discussed in detail:

1. Sampling and Storage Sampling methods and program. Contamination of the sample during sampling and storage. Changes in the composition during storage. Various storage methods for solid and liquid samples.
2. Sample Preparation Drying and ashing. Wet digestion. Different methods for preparing samples of hard, semihard, and soft tissues, and of biological fluids.

     

Study of biological materials based on indentometry

The general multiparameter system of testing chemical compounds allows to study biological matter on the basis of purely physical approaches. A "Tissue-1" device allows to study all types of biological tissues able to withstand different kinds of passive and/or active mechanic load.     

Rapid prototyping in tissue engineering: challenges and potential

Tissue engineering aims to produce patient-specific biological substitutes in an attempt to circumvent the limitations of existing clinical treatments for damaged tissue or organs. The main regenerative tissue engineering approach involves transplantation of cells onto scaffolds. The scaffold attempts to mimic the function of the natural extracellular matrix, providing a temporary template for the growth of target tissues. Scaffolds should have suitable architecture and strength to serve their intended function. This paper presents a comprehensive review of the fabrication methods, including conventional, mainly manual, techniques and advanced processing methods such as rapid prototyping (RP) techniques. The potential and challenges of scaffold-based technology are discussed from the perspective of RP technology.     

Imaging Brain Activity With Voltage- and Calcium-Sensitive Dyes

This paper presents three examples of imaging brain activity with voltage- or calcium-sensitive dyes and then discusses the methodological aspects of the measurements that are needed to achieve an optimal signal-to-noise ratio.Internally injected voltage-sensitive dye can be used to monitor membrane potential in the dendrites of invertebrate and vertebrate neurons in in vitro preparations.Both invertebrate and vertebrate ganglia can be bathed in voltage-sensitive dyes to stain all of the cell bodies in the preparation. These dyes can then be used to follow the spike activity of many neurons simultaneously while the preparations are generating behaviors.Calcium-sensitive dyes that are internalized into olfactory receptor neurons in the nose will, after several days, be transported to the nerve terminals of these cells in the olfactory bulb. There they can be used to measure the input from the nose to the bulb.Three kinds of noise are discussed. a. Shot noise from the random emission of photons from the preparation. b. Vibrational noise from external sources. c. Noise that occurs in the absence of light, the dark noise.Three different parts of the light measuring apparatus are discussed: the light sources, the optics, and the cameras.The major effort presently underway to improve the usefulness of optical recordings of brain activity are to find methods for staining individual cell types in the brain. Most of these efforts center around fluorescent protein sensors of activity.     

Evaluation of Small Intestine Grafts Decellularization Methods for Corneal Tissue Engineering

Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.     

High-resolution confocal imaging and three-dimensional rendering

Intrinsic opacity and inhomeogeniety of most biological tissues have prevented the efficient light penetration and signal detection for high-resolution confocal imaging of thick tissues. Here, we summarize recent technical advances in high-resolution confocal imaging for visualization of cellular structures and gene expression within intact whole-mount thick tissues. First, we introduce features of the FocusClear technology that render biological tissue transparent and thus improve the light penetration and signal detection. Next, a universal fluorescence staining method that labels all nuclei and membranes is described. We then demonstrate the postrecording image processing techniques for 3D visualization. From these images, regions of interest in the whole-mount brain can be segmented and volume rendered. Together, these technical advances in confocal microscopy allow visualization of structures within whole-mount tissues up to 1mm thick at a resolution similar to that of the observation of single cells in culture. Practical uses and limitations of these techniques are discussed.