Laser Capture Microdissection Coupled with On-Column Extraction LC-MSn Enables Lipidomics of Fluorescently Labeled Drosophila Neurons
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- 作者：Sarita Hebbar ; Wolf Dieter Schulz ; Ulrich Sauer ; Dominik Schwudke
- 刊名：Analytical Chemistry
- 出版年：2014
- 出版时间：June 3, 2014
- 年：2014
- 卷：86
- 期：11
- 页码：5345-5352
- 全文大小：484K
- 年卷期：v.86,no.11(June 3, 2014)
- ISSN：1520-6882
文摘
We have used laser capture microdissection (LCM) and fluorescence microscopy to isolate genetically labeled neurons from the Drosophila melanogaster brain. From native thin sections, regions of interest could be analyzed with a spatial resolution better than 50 渭m. To exploit the specificity of LCM for lipidomics, catapulted tissue patches were directly collected on a reversed phase column and analyzed using an on-column extraction (OCE) that was directly coupled with liquid chromatography鈥搈ultistage mass spectrometry (LC-MSn). With this approach, more than 50 membrane lipids belonging to 9 classes were quantified in tissue regions equivalent to a sample amount of 50 cells. Using this method, the limit of quantitation and the extraction efficiency could be estimated enabling a reliable evaluation of acquired lipid profiles. The lipid profiles of cell body- and synapse-enriched regions of the Drosophila brain were determined and found to be distinct. We argue that this workflow represents a tremendous improvement for tissue lipidomics by integrating genetics, fluorescence microscopy, LCM and LC-MSn.