Quanico Jusal, Franck Julien, Wisztorski Maxence, Salzet Michel, Fournier Isabelle
Université de Lille 1, Inserm, U-1192 - Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse (PRISM), F-59000, Lille, France.
Methods Mol Biol. 2017;1598:269-283. doi: 10.1007/978-1-4939-6952-4_13.
Proteins and other biomolecules such as lipids are significant players in the central nervous system and are implicated in various neurological disorders. Their identification, quantification, and distribution are thus important not only in understanding the disease but also in developing treatments. A combined workflow allowing the localized microextraction of discrete regions identified by a matrix-assisted laser desorption/ionization mass spectrometry (MSI) imaging experiment for proteomics analysis by liquid chromatography/tandem mass spectrometry (LC MS/MS) is described in this chapter. MSI was initially used to map lipid distributions allowing for the identification of regions of interest (ROIs) that are then subjected to microextraction in a consecutive tissue section. Mounting of consecutive tissue on parafilm allows microdissection of the ROIs, where proteins can then be recovered for processing and LC MS/MS analysis. The PAM method provides a fast and cheap means to perform further downstream analysis after an MSI experiment.
蛋白质和其他生物分子(如脂质)在中枢神经系统中起着重要作用,并与各种神经系统疾病有关。因此,它们的鉴定、定量和分布不仅对于理解疾病很重要,而且对于开发治疗方法也很重要。本章介绍了一种组合工作流程,该流程允许对通过基质辅助激光解吸/电离质谱(MSI)成像实验确定的离散区域进行局部微萃取,以便通过液相色谱/串联质谱(LC MS/MS)进行蛋白质组学分析。MSI最初用于绘制脂质分布图,从而识别感兴趣的区域(ROI),然后在连续的组织切片中对这些区域进行微萃取。将连续的组织固定在石蜡膜上可以对ROI进行显微切割,然后可以回收蛋白质进行处理和LC MS/MS分析。PAM方法提供了一种在MSI实验后进行进一步下游分析的快速且廉价的手段。