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利用傅里叶变换红外显微光谱法对正常和恶性前列腺组织进行表征

Characterization of normal and malignant prostate tissue by Fourier transform infrared microspectroscopy.

作者信息

Pezzei Christine, Pallua Johannes D, Schaefer Georg, Seifarth Christof, Huck-Pezzei Verena, Bittner Lukas K, Klocker Helmut, Bartsch Georg, Bonn Guenther K, Huck Christian W

机构信息

Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 52a, 6020 Innsbruck, Austria.

出版信息

Mol Biosyst. 2010 Nov;6(11):2287-95. doi: 10.1039/c0mb00041h. Epub 2010 Sep 27.

Abstract

Prostate cancer has become one of the most common malignancies worldwide. Morphological and histomorphological evaluation of this disease is a well established technique for the cancer classification and has remained relatively unchanged since several decades, although it remains a time consuming and subjective technique, with unsatisfactory levels of inter- and intra-observer discrepancy. Novel approaches for histological recognition are necessary to identify and to investigate cancer in detail. Fourier transform infrared (FTIR) spectroscopic imaging has become an essential tool for the detection, identification and characterization of the molecular components of biological processes, such as those responsible for the dynamic properties of cancer progression. Major advantage of this new technique is the acquisition of local molecular expression profiles while maintaining the topographic integrity of the tissue and avoiding time-consuming extraction, purification and separation steps. By using this method it is possible to investigate the spatial distribution of proteins, lipids, carbohydrates, cholesterols, nucleic acids, phospholipids and small molecules within biological systems by in situ analysis of tissue sections. We applied this technique on prostate cancer patients radical prostatectomy specimens in order to develop new tools for histomorphological analysis and the characterization of snap frozen prostate cancer tissues. As a first step, an optimization of sample preparation, tissue section thickness and IR slide material was performed. Special preparation methods for FTIR imaging are the essential requirements to maintain the spatial arrangement of compounds and avoid delocalization and degradation of the analytes. Subsequently, selected cancer samples were characterized with the prior optimized parameters and analyzed by univariate and cluster analysis. For the interpretation and calibration of the system we correlated the FTIR-images with the histopathological information. With this method it is possible to distinguish between cancer and noncancer areas within a prostate cancer tissue with a resolution of 6.25 μm × 6.25 μm on frozen sections.

摘要

前列腺癌已成为全球最常见的恶性肿瘤之一。对这种疾病进行形态学和组织形态学评估是一种成熟的癌症分类技术,几十年来相对没有变化,尽管它仍然是一种耗时且主观的技术,观察者间和观察者内的差异水平不尽人意。需要新的组织学识别方法来详细识别和研究癌症。傅里叶变换红外(FTIR)光谱成像已成为检测、识别和表征生物过程分子成分的重要工具,例如那些负责癌症进展动态特性的成分。这项新技术的主要优点是在保持组织地形完整性的同时获取局部分子表达谱,并且避免了耗时的提取、纯化和分离步骤。通过使用这种方法,可以通过对组织切片进行原位分析来研究生物系统中蛋白质、脂质、碳水化合物、胆固醇、核酸、磷脂和小分子的空间分布。我们将这项技术应用于前列腺癌患者的根治性前列腺切除术标本,以开发用于组织形态学分析和快速冷冻前列腺癌组织表征的新工具。第一步,对样品制备、组织切片厚度和红外载玻片材料进行了优化。FTIR成像的特殊制备方法是保持化合物空间排列并避免分析物离域和降解的基本要求。随后,使用先前优化的参数对选定的癌症样本进行表征,并通过单变量和聚类分析进行分析。为了对系统进行解释和校准,我们将FTIR图像与组织病理学信息相关联。通过这种方法,可以在冷冻切片上以6.25μm×6.25μm的分辨率区分前列腺癌组织内的癌区和非癌区。

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