School of Engineering, University of Portsmouth, Portsmouth, UK.
School of Pharmacy and Institute of Biomedical and Biomolecular Science, University of Portsmouth, Portsmouth, UK.
J Microsc. 2018 Dec;272(3):180-195. doi: 10.1111/jmi.12719. Epub 2018 Jun 6.
Fibrous nanomaterials such as electrospun materials have many uses ranging from tissue engineering to biosensors. High-resolution imaging is an important component in the characterization of these materials. Important parameters required to predict and study the properties of fibre rich materials include diameter and orientation distribution as well as fibre spacing. The orientations and the relative dimensions of the fibres can be measured via specially designed imaging software. Difficulties in this measurement process can arise if fibres are distributed in close proximity to each other in relation to the resolution of the imaging modality. For example, if some automation is required in the measurement process and, particularly, if the automated processes are not designed for situations where the fibres are in close proximity to each other. This work is therefore concerned with the development of automated measurement techniques to provide estimates of the diameters of fibres and also the orientation distribution. The software automatically detects special points in the fibrous materials where fibres can be considered to have some delineation from surrounding fibres. These sparse points are considered to be points at which estimates of the fibres' properties can be quantified. Aligned and randomly distributed electrospun poly(caprolactone) nanofibres were prepared. Imaging of these materials was performed with an X-ray Computer Tomography system with an image voxel size of 0.15 × 0.15 × 0.15 μm . Scanning Electron Microscopy images were also obtained. Fibre diameters estimated using images from both modalities using the developed techniques were found to be in agreement. Orientation distribution was summarized with multiscale entropy and found to be consistent with visual observation across different scales. LAY DESCRIPTION: Fibres are present in many types of materials which can include, for some materials, very small fibres e.g. a few nanometres in diameter. Very small fibres are present in collagen and elastin which are common tissues of many organs in many types of living things. The sizes of these very small fibres and how they are arranged are important information that can help in the understanding of the overall properties of these materials. Materials with very small fibres can also be synthesized using specialised techniques. The properties of these synthesized fibrous materials are also important to help in understanding how the materials will perform in various different applications. Applications are many and can range from tissue engineering to drug delivery. Some properties of these materials can be shown, visually, with the aid of 3D imaging techniques such as X-ray Computer Tomography (XCT) or in 2D, with Scanning Electron Microscopy (SEM) but at a higher magnification. The work described here is centred around the development of computer algorithms to automatically determine material properties from 3D XCT images. Tests are performed with material samples, where the fibres are aligned (in semi-parallel fashion) and another where the fibres are randomly oriented (criss-crossing). The tests show that the developed algorithms are able to successfully and relatively accurately determine the diameters of the fibres. The tests also show that it is possible to quantify the relative randomness of the orientations of the fibres.
纤维状纳米材料,如静电纺丝材料,具有许多用途,从组织工程到生物传感器都有涉及。高分辨率成像是这些材料特性表征的一个重要组成部分。预测和研究富含纤维材料特性所需的重要参数包括直径和取向分布以及纤维间距。纤维的取向和相对尺寸可以通过专门设计的成像软件进行测量。如果纤维彼此之间的分布非常接近,以至于成像方式的分辨率无法分辨,那么在这个测量过程中就会出现困难。例如,如果测量过程需要自动化,特别是如果自动化过程不是为纤维彼此非常接近的情况设计的。因此,这项工作致力于开发自动化测量技术,以提供纤维直径和取向分布的估计值。软件会自动检测纤维材料中的特殊点,这些点可以认为是纤维与周围纤维有一些区别的地方。这些稀疏的点被认为是可以量化纤维特性的估计点。本研究制备了取向和随机分布的静电纺聚己内酯纳米纤维。使用具有 0.15×0.15×0.15 μm 图像体素大小的 X 射线计算机断层扫描系统对这些材料进行成像。还获得了扫描电子显微镜图像。使用开发的技术从两种模式的图像中估计纤维直径,结果发现它们是一致的。使用多尺度熵对取向分布进行总结,发现与不同尺度的视觉观察结果一致。