DEVELOPMENT OF FAR INFRARED MICROSPECTROSCOPY AT BEAMLINE U2B

光束线 U2B 远红外显微光谱学的发展

• 批准号: 6345106
• 负责人: LISA M MILLER
• 金额: $ 2.8万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6345106
• 关键词: aging; animal tissue; biomedical resource; spectrometry

A standard method for preserving bone tissue samples is to embed them in paraffin or plastic (methyl methacrylate/dibutyl phthalate). Then they can be sectioned at 5-10 ?m thicknesses for transmission infrared experiments. These thin sections are required due to the extremely large signals provided by the phosphate and protein contributions to the infrared spectra. Although embedding provides an excellent means for examining the mineral components of bone, this technique has a few drawbacks including (1) interference of the infrared absorbance of the embedding medium, and (2) it is uncertain whether the embedding process affects protein secondary structure. Conventional freeze/microtome cleavage methods can be used to provide native tissue samples. However, it is difficult to section to less than 20 ?m, which is problematic due to the high infrared absorbance at those thicknesses. Thus, we are using attenuated total reflection infrared microspectroscopy to examine such native samples (eg. without plastic). Sections of tissue are frozen and cleaved with a microtome and examined using Attenuated Total Reflectance (ATR). ATR is a surface technique that probes the tissue structure approximately 10 ?m into the surface based on the evanescent wavefront of the infrared radiation. Due to this property, the samples are examined wet. ATR infrared microspectroscopy is a superior method for examining protein spectra, since no denaturing conditions that may perturb the protein structure are required.

保存骨组织样本的标准方法是嵌入 它们在石蜡或塑料（甲基丙烯酸甲酯/邻苯二甲酸二丁酯）中。 然后可以将它们切成 5-10 µm 厚度的切片以进行传输 红外实验。 需要这些薄片是因为 磷酸盐和蛋白质提供极大的信号 对红外光谱的贡献。 虽然嵌入提供了 这是检查骨骼矿物质成分的绝佳方法 该技术有一些缺点，包括（1） 包埋介质的红外吸光度，以及 (2) 不确定 嵌入过程是否影响蛋白质二级结构。 传统的冷冻/切片机裂解方法可用于提供 天然组织样本。 然而，很难分割到更少 小于 20 µm，由于红外吸收率高，这是有问题的 在那些厚度下。 因此，我们使用衰减全反射 红外显微光谱法来检查此类天然样品（例如。 无塑料）。 组织切片被冷冻并用 切片机并使用衰减全反射（ATR）进行检查。 衰减全反射 是一种表面技术，可大致探测组织结构 基于渐逝波前的表面 10 微米 红外线辐射。 由于这一特性，对样品进行了检查 湿。 ATR 红外显微光谱法是一种优越的方法 检查蛋白质光谱，因为没有可能的变性条件 扰乱蛋白质结构是必需的。