期刊
PROTEOMICS
卷 16, 期 11-12, 页码 1652-1659出版社
WILEY
DOI: 10.1002/pmic.201500402
关键词
Funnel-freezing; Heat-stabilization; Ischemic stroke; MALDI-MSI; Metabolites; Technology
资金
- ZonMW Zenith project Imaging Mass Spectrometry-Based Molecular Histology: Differentiation and Characterization of Clinically Challenging Soft Tissue Sarcomas [93512002]
- Dutch Heart Foundation [2011T055]
- Dutch Brain Foundation [2011(1)-102]
- ZonMW Veni grant
- Centre for Medical Systems Biology (CMSB) in the framework of the Netherlands Genomics Initiative (NGI)
- Marie Curie Career Integration Grant [294233]
- FP7 EUROHEADPAIN [602633]
- Marie Curie IAPP Program BRAINPATH [612360]
Tissue preparation is the key to a successful matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) experiment. Rapid post-mortem changes contribute a significant challenge to the use of MSI approaches for the analysis of peptides and metabolites. In this technical note we aimed to compare the tissue fixation method ex-vivo heat-stabilization with in-situ funnel-freezing in a middle cerebral artery occlusion (MCAo) mouse model of stroke, which causes profound alterations in metabolite concentrations. The influence of the duration of the thaw-mounting of the tissue sections on metabolite stability was also determined. We demonstrate improved stability and biomolecule visualization when funnel-freezing was used to sacrifice the mouse compared with heat-stabilization. Results were further improved when funnel-freezing was combined with fast thaw-mounting of the brain sections.
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