Journal
ANALYTICAL CHEMISTRY
Volume 86, Issue 11, Pages 5345-5352Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac500276r
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Funding
- Wellcome Trust/DBT India
- NCBS-Merck Co
- NCBS-TIFR
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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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