Funnel-freezing versus heat-stabilization for the visualization of metabolites by mass spectrometry imaging in a mouse stroke model

被引：23
作者：
Mulder, Inge A. [1 ]
Esteve, Clara [2 ]
Wermer, Marieke J. H. [1 ]
Hoehn, Mathias [3 ,4 ,5 ]
Tolner, Else A. [1 ,6 ]
van den Maagdenberg, Arn M. J. M. [1 ,6 ]
McDonnell, Liam A. [2 ,7 ]
机构：
[1] Leiden Univ, Dept Neurol, Med Ctr, Leiden, Netherlands
[2] Leiden Univ, Ctr Prote & Metabol, Med Ctr, Einthovenweg 20, NL-2333 ZC Leiden, Netherlands
[3] Leiden Univ, Med Ctr, Dept Radiol, Leiden, Netherlands
[4] Percuros BV, Enschede, Netherlands
[5] Max Planck Inst Neurol Res, In Vivo NMR Lab, Cologne, Germany
[6] Leiden Univ, Med Ctr, Dept Human Genet, Leiden, Netherlands
[7] Fdn Pisana Sci ONLUS, Pisa, Italy
来源：
PROTEOMICS | 2016年 / 16卷 / 11-12期
关键词：
Funnel-freezing; Heat-stabilization; Ischemic stroke; MALDI-MSI; Metabolites; Technology; CEREBRAL METABOLITES; TISSUES; RATS; LOCALIZATION; TEMPERATURE; SECTIONS; ISCHEMIA; PENUMBRA; STATE;
D O I：
10.1002/pmic.201500402
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：1652 / 1659
页数：8
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