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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: MPI-CBG Publications 2014 (arch)     Display Documents

ID: 705669.0, MPI für molekulare Zellbiologie und Genetik / MPI-CBG Publications 2014 (arch)
Laser capture microdissection coupled with on-column extraction LC-MS(n) enables lipidomics of fluorescently labeled Drosophila neurons.
Authors:Hebbar, Sarita; Schulz, Wolf Dieter; Sauer, Ulrich; Schwudke, Dominik
Date of Publication (YYYY-MM-DD):2014
Title of Journal:Analytical Chemistry
Issue / Number:11
Start Page:5345
End Page:5352
Copyright:not available
Audience:Experts Only
Intended Educational Use:No
Abstract / Description: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).
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:thuem
Affiliations:MPI für molekulare Zellbiologie und Genetik
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