Ami Diletta, Natalello Antonino, Doglia Silvia Maria
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.
Methods Mol Biol. 2012;895:85-100. doi: 10.1007/978-1-61779-927-3_7.
Fourier transform infrared (FTIR) microspectroscopy is a powerful tool for the study of complex biological systems. Indeed, it is employed to characterize intact cells, tissues, and whole model organisms such as nematodes, since it allows to obtain a chemical fingerprint of the sample under investigation, giving information on the molecular composition and structures. The successful application of this technique for the in situ study of biological processes requires specific sample preparations, in order to obtain reliable and reproducible results. In the present work, we illustrate the optimized procedures to prepare biological samples for IR measurements and the method to collect and analyze their FTIR spectra. In particular, we describe here the investigations on bacterial cells, intact eukaryotic cells, and whole intact nematode specimens.
傅里叶变换红外(FTIR)显微光谱学是研究复杂生物系统的有力工具。实际上,它被用于表征完整的细胞、组织以及诸如线虫等整个模式生物,因为它能够获取被研究样品的化学指纹图谱,提供有关分子组成和结构的信息。要成功应用该技术进行生物过程的原位研究,需要特定的样品制备方法,以便获得可靠且可重复的结果。在本工作中,我们阐述了用于红外测量的生物样品制备的优化程序以及收集和分析其FTIR光谱的方法。特别是,我们在此描述了对细菌细胞、完整真核细胞和完整线虫标本的研究。