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Analyses of biological systems using scanning near-field infrared microscopy.

机译：使用扫描近场红外显微镜对生物系统进行分析。

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摘要

We report the construction of a Scanning Near-field Infrared Microscope (SNIM) and its successful operation towards infrared microspectroscopic analysis of biological tissue. Infrared microspectroscopy is a useful technique that can rapidly identify and localize the chemical constituents within biological tissues and cells on the basis of the vibrational spectroscopic signatures of their organic functional groups. Such analysis may be particularly useful if carried out using near-field optics so as to achieve subwavelength resolution unavailable to Fourier Transform InfraRed (FTIR) microscopy. We constructed SNIM by adapting a conventional scanning near-field microscope for use in the infrared. In particular, we employ infrared-transmitting chalcogenide fibers to serve as the fiber optic probes, and a free electron laser, a laser tunable in the mid-infrared wavelength range, is used as the illumination source. SNIM has acquired mid-infrared images of metal, semiconductor, and biological samples. We have demonstrated resolution down to 2.5 micrometers, and the device appears to be sensitive to differences in sample absorption of less than 0.1 OD. We used SNIM to examine unstained microtome sections of human atherosclerotic lesions, both by infrared imaging and by localized vibrational spectroscopy. In this way, we were able to identify and localize protein, lipid, and mineral components within the tissue. The studies of atherosclerotic tissue illustrate the usefulness of SNIM towards in situ vibrational microspectroscopic investigation of biological systems. In addition, SNIM has demonstrated the ability to perform infrared microscopic imaging in a liquid medium, and we have successfully used the device to take images of living cells in a liquid environment. We believe that SNIM provides a unique opportunity to study the cellular processes of living cells and bacteria by spectroscopic means.
机译：我们报告的扫描近场红外显微镜（SNIM）的建设及其对生物组织的红外光谱分析的成功运作。红外显微技术是一种有用的技术，可以根据其有机官能团的振动光谱特征快速识别和定位生物组织和细胞内的化学成分。如果使用近场光学器件进行这种分析，以获得傅里叶变换红外（FTIR）显微镜无法获得的亚波长分辨率，则这种分析可能特别有用。我们通过将常规扫描近场显微镜用于红外来构建SNIM。尤其是，我们采用红外透射硫族化物纤维作为光纤探针，而自由电子激光器（可在中红外波长范围内可调的激光器）用作照明源。 SNIM已获取金属，半导体和生物样品的中红外图像。我们已经证明分辨率低至2.5微米，并且该设备似乎对小于0.1 OD的样品吸收差异敏感。我们使用SNIM通过红外成像和局部振动光谱检查了人类动脉粥样硬化病变的未染色切片机切片。通过这种方式，我们能够识别并定位组织中的蛋白质，脂质和矿物质成分。动脉粥样硬化组织的研究表明，SNIM对于生物系统的原位振动显微光谱研究很有用。此外，SNIM展示了在液体介质中执行红外显微成像的能力，并且我们已经成功地使用该设备在液体环境中拍摄活细胞的图像。我们相信，SNIM提供了独特的机会，可以通过光谱学方法研究活细胞和细菌的细胞过程。

著录项

  • 作者

    Jeung, Andrew G.;

  • 作者单位

    Stanford University.;

  • 授予单位 Stanford University.;
  • 学科 Physics Optics.; Biophysics Medical.
  • 学位 Ph.D.
  • 年度 1998
  • 页码 113 p.
  • 总页数 113
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 光学;生物物理学;
  • 关键词

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