期刊
ANALYTICAL CHEMISTRY
卷 86, 期 11, 页码 5345-5352出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac500276r
关键词
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资金
- Wellcome Trust/DBT India
- NCBS-Merck Co
- NCBS-TIFR
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 mu 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-multistage mass spectrometry (LC-MSn). With this approach, more than SO membrane lipids belonging to 9 classes were quantified in tissue regions equivalent to a sample amount of SO 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 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.
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