当前位置: X-MOL 学术Anal. Methods › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Empirical study on the effects of acquisition parameters for FTIR hyperspectral imaging of brain tissue.
Analytical Methods ( IF 3.1 ) Pub Date : 2020-08-11 , DOI: 10.1039/c9ay01200a
J Sacharz 1 , D Perez-Guaita 2 , Mustafa Kansiz 3 , Shaiju S Nazeer 4 , A Wesełucha-Birczyńska 5 , S Petratos 6 , B R Wood 4 , P Heraud 7
Affiliation  

Fourier transform infrared (FTIR) spectroscopic imaging is a powerful technique for molecular imaging of pathologies associated with the nervous systems including multiple sclerosis research. However, there is no standard methodology or standardized protocol for FTIR imaging of tissue sections that maximize the ability to discriminate between the molecular, white and granular layers, which is essential in the investigation of the mechanism of demyelination process. Tissue sections are heterogeneous, complex and delicate, hence the parameters to generate high quality images in minimal time becomes essential in the modern clinical laboratory. This article presents an FTIR spectroscopic imaging study of post-mortem human brain tissue testing the effects of various measurement parameters and data analysis methods on image quality and acquisition time. Hyperspectral images acquired from the same region of a tissue using a range of the most common optical and collection parameters in different combinations were compared. These included magnification (4× and 15×), number of co-added scans (1, 4, 8, 16, 32, 64 and 128 scans) and spectral resolution (4, 8 and 16 cm−1). Images were compared in terms of acquisition time, signal-to-noise (S/N) ratio, and accuracy of the discrimination between three major tissue types in a section from the cerebellum (white matter, granular and molecular layers). In the latter case, unsupervised k-means cluster (KMC) analysis was employed to generate images from the hyperspectral images, which were compared to a reference image. The classification accuracy for tissue class discrimination was highest for the 4× magnifying objective, with 4 cm−1 spectral resolution and 128 co-added scans. The 15× magnifying objective gave the best accuracy for a spectral resolution of 4 cm−1 and 64 scans (96.3%), which was just above what was achieved using the 4× magnifying objective, with 4 cm−1 spectral resolution and 32 and 64 co-added scans (95.4 and 95.6%, respectively). These findings were correlated with a decrease in S/N ratio with increasing number of scans and was generally lower for the 15× objective. However, longer scan times were required using the 15× magnifying objective, which did not justify the very small improvement in the classification of tissue types.

中文翻译:

采集参数对脑组织FTIR高光谱成像影响的实证研究。

傅里叶变换红外(FTIR)光谱成像技术是对与神经系统有关的病理学进行分子成像的一项强大技术,包括多发性硬化症研究。但是,没有用于组织切片的FTIR成像的标准方法或标准化协议,该方法可以最大程度地区分分子层,白色层和颗粒层,这在研究脱髓鞘过程的机制中至关重要。组织切片是异质,复杂和脆弱的,因此在现代临床实验室中,在最短时间内生成高质量图像的参数就变得至关重要。本文介绍了人体后组织验尸的FTIR光谱成像研究,测试了各种测量参数和数据分析方法对图像质量和采集时间的影响。比较了使用不同组合的一系列最常见的光学和采集参数从组织的同一区域获取的高光谱图像。其中包括放大倍数(4倍和15倍),共加扫描的次数(1、4、8、16、32、64和128扫描)和光谱分辨率(4、8和16厘米)-1)。比较图像的采集时间,信噪比(S / N)和小脑切片中三种主要组织类型(白质,颗粒和分子层)的区分精度。在后一种情况下,采用无监督k均值聚类(KMC)分析从高光谱图像生成图像,并将其与参考图像进行比较。对于4倍放大物镜,组织类别识别的分类精度最高,具有4 cm -1的光谱分辨率和128次共加扫描。15倍物镜在4 cm -1的光谱分辨率和64次扫描(96.3%)时提供了最佳的精度,刚好高于4倍物镜的4 cm扫描分辨率-1光谱分辨率和32和64共加扫描(分别为95.4和95.6%)。这些发现与信噪比随扫描次数增加而降低有关,通常对于15倍物镜而言更低。但是,使用15倍物镜需要更长的扫描时间,这不能证明组织类型的分类有很小的改善。
更新日期:2020-09-17
down
wechat
bug