Label-free Raman spectroscopic imaging to extract morphological and chemical information from a formalin-fixed, paraffin-embedded rat colon tissue section.
Label-free Raman spectroscopic imaging to extract morphological and chemical information from a formalin-fixed, paraffin-embedded rat colon tissue section.
复制标题
DOI：
10.1111/iep.12194
复制
发表时间：
2016-08
影响因子：
3
通讯作者：
Thomas GM
中科院分区：
文献类型：
--
作者：
Gaifulina R;Maher AT;Kendall C;Nelson J;Rodriguez-Justo M;Lau K;Thomas GM
动物模型和存档的人类生物库组织是疾病发展、诊断和治疗研究的有用资源。为了保存显微解剖特征并便于长期储存，大多数组织样本通过固定、石蜡包埋和后续脱蜡所需的化学处理进行变性。这些侵蚀性的化学过程被认为会改变样品的生化组成，并可能损害可用于诊断或测序的可靠光谱检查。因此，光谱分析通常在新鲜/冷冻样品上进行。在这项研究中，我们提供了一个广泛的生化信号处理后的样品（福尔马林固定和石蜡包埋，FFPE），特别是那些来自健康大鼠结肠的解剖层的表征。化学计量学分析方法（无监督和监督）的应用表明，消除了对组织染色的需要，并很容易揭示与杯状细胞和粘膜内密集细胞群一致的显微特征，主要是通过强核酸信号。我们还能够识别胶原粘膜下层和浆膜，以及来自肌肉区域和血管的肌肉相关信号。对数据应用线性回归分析，我们能够通过参考真实生物分子标准品的子集确认每层的生物来源来证实细胞和组织类型的这种初始分配。我们的研究结果表明，直接与常规苏木精和伊红（H & E）染色相比，使用无标记拉曼显微光谱在FFPE切片中获得上级成像对比度的潜力。
Animal models and archived human biobank tissues are useful resources for research in disease development, diagnostics and therapeutics. For the preservation of microscopic anatomical features and to facilitate long‐term storage, a majority of tissue samples are denatured by the chemical treatments required for fixation, paraffin embedding and subsequent deparaffinization. These aggressive chemical processes are thought to modify the biochemical composition of the sample and potentially compromise reliable spectroscopic examination useful for the diagnosis or biomarking. As a result, spectroscopy is often conducted on fresh/frozen samples. In this study, we provide an extensive characterization of the biochemical signals remaining in processed samples (formalin fixation and paraffin embedding, FFPE) and especially those originating from the anatomical layers of a healthy rat colon. The application of chemometric analytical methods (unsupervised and supervised) was shown to eliminate the need for tissue staining and easily revealed microscopic features consistent with goblet cells and the dense populations of cells within the mucosa, principally via strong nucleic acid signals. We were also able to identify the collagenous submucosa‐ and serosa‐ as well as the muscle‐associated signals from the muscular regions and blood vessels. Applying linear regression analysis to the data, we were able to corroborate this initial assignment of cell and tissue types by confirming the biological origin of each layer by reference to a subset of authentic biomolecular standards. Our results demonstrate the potential of using label‐free Raman microspectroscopy to obtain superior imaging contrast in FFPE sections when compared directly to conventional haematoxylin and eosin (H&E) staining.
登录
查看更多内容
影响因子：
3.4
作者：
Klein, Katharina;Gigler, Alexander M.;Schlegel, Juergen
通讯作者：
Schlegel, Juergen
影响因子：
--
作者：
Chowdary, M. V. P.;Kumar, K. Kalyan;Mathew, Stanley
通讯作者：
Mathew, Stanley
影响因子：
3.1
作者：
CASSIDY, MM;LIGHTFOOT, FG;VAHOUNY, GV
通讯作者：
VAHOUNY, GV
影响因子：
78.5
作者：
Kufe, Donald W.
通讯作者：
Kufe, Donald W.
影响因子：
6.6
作者：
De Jong, BWD;Schut, TCB;Puppels, GJ
通讯作者：
Puppels, GJ