title: Laser capture microdissection coupled with on-column extraction LC-MS(n) enables lipidomics of fluorescently labeled Drosophila neurons.
abstract: 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 μm. 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-MS(n)). 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-MS(n).
authors.affiliation: National Centre for Biological Sciences, Tata Institute of Fundamental Research , Bangalore 560065, India.
authors.forename: Sarita
authors.initials: S
authors.lastname: Hebbar
authors.forename: Wolf Dieter
authors.initials: WD
authors.lastname: Schulz
authors.forename: Ulrich
authors.initials: U
authors.lastname: Sauer
authors.forename: Dominik
authors.initials: D
authors.lastname: Schwudke
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004331:Drosophila melanogaster / Q000737:chemistry; D061346:Laser Capture Microdissection / Q000379:methods*; D007834:Lasers; D008055:Lipids / Q000737:chemistry*; D013058:Mass Spectrometry; D009474:Neurons / Q000737:chemistry*
chemical_list: D008055:Lipids