Mid-FTIR-Microspectroscopy of stratum corneum single cells and stratum corneum tissue
Mid-FTIR-Microspectroscopy of stratum corneum single cells and stratum corneum tissue
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DOI：
10.1039/b207478h
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发表时间：
2002-11-15
影响因子：
3.3
通讯作者：
Garidel, P
中科院分区：
文献类型：
--
作者：
Garidel, P
角质层(SC)是皮肤最上层的组织。它保护身体免受外界冲击，防止身体干燥。角质层是一种薄组织，大小约20微米，主要由两部分组成：脂类和蛋白质。蛋白质位于角膜细胞中，这些细胞嵌入连续的脂质基质中。角质层尚未通过横向空间分辨率为20x20um的红外光谱进行详细检查，这是在单个细胞水平上的分辨率所必需的。本研究报告了猪干细胞和单个角质细胞的中傅里叶变换红外显微光谱分析(M-FTIR-MSP)，目的是确定影响其红外光谱的特定分子成分。与基于标准光学显微镜的程序相比，红外方法既不需要耗时的样本染色，也不需要固定。此外，还利用衰减全反射傅里叶变换红外光谱(ATR-FTIR)对不同的SC组分进行了分析，识别出了特征的生物标志物条带。从单个角质细胞获得的M-FTIR-MSP光谱中，脂肪组分可以很容易地通过较强的亚甲基伸缩振动以及羰基酯带来识别。酰胺I和II以及对称的甲基吸收峰清楚地归属于蛋白质组分，可用于组织蛋白质的鉴定。此外，角质细胞分数还可以通过位于3070厘米(-1)附近的谱带来表征，这归因于酰胺B的振动。对红外数据的分析表明，红外光谱分析方法对组织生化的变化以及SC组织结构和组织的变化具有很高的敏感性。这项研究是今后旨在开发红外显微光谱成像技术作为组织特征的补充诊断工具的研究的先决条件。
The stratum corneum (SC) is the uppermost tissue of the skin. It protects the body against external impacts and prevents it from desiccation. The stratum corneum is a thin tissue of similar to20 mum, which is mainly composed of two fractions: lipids and proteins. The proteins are located in the corneocytes and these cells are embedded in a continuous lipid matrix.Stratum corneum has not yet been examined in detail by infrared spectroscopy at a lateral spatial resolution of 20 x 20 mum, which is required for resolutions at the level of individual cells. The current study reports mid Fourier transform infrared microspectroscopic (M-FTIR-MSP) analyses of porcine SC and single corneocyte cells with the aim to identify the specific molecular components that contribute to its infrared spectrum. In contrast to standard light microscopy based procedures, the infrared approach requires neither time consuming sample staining nor fixation. It is therefore, a non-invasive and rapid bio-analytical method.In addition to this, the different SC fractions were also analysed by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy.Characteristic biomarker bands have been recognized. The lipid fraction can easily be identified by the strong methylene stretching vibrations as well as by the carbonyl ester band, which were absent in the M-FTIR-MSP spectra obtained from single corneocytes. The amide I and II as Well as the symmetric methyl absorptions are clearly assigned to the protein fraction and can be used for tissue protein identification. In addition to this, the corneocyte fraction can also be characterised by a band located around 3070 cm(-1), which is attributed to the amide B vibration.The analysis of the IR data reveals a high sensitivity of the infrared spectroscopy approach toward changes in tissue biochemistry and variation in SC tissue architecture and organisation. This study is a prerequisite for future studies aimed at the development of infrared microspectroscopic imaging techniques as a complementing diagnostic tool for the characterisation of tissues.