High-mass-resolution MALDI mass spectrometry imaging of metabolites from formalin-fixed paraffin-embedded tissue
High-mass-resolution MALDI mass spectrometry imaging of metabolites from formalin-fixed paraffin-embedded tissue
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DOI：
10.1038/nprot.2016.081
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发表时间：
2016-08-01
期刊：
影响因子：
14.8
通讯作者：
Walch, Axel
中科院分区：
文献类型：
--
作者：
Ly, Alice;Buck, Achim;Walch, Axel
福尔马林固定和石蜡包埋(FFPEPE)组织标本是组织学检查的金标准，它们在组织研究中提供了有价值的分子信息。由于缺乏适当的程序，以及对组织处理过程中代谢物含量或化学状态变化的担忧，对存档组织样本的代谢物评估尚未广泛进行。我们提出了一种使用高质量分辨率基质辅助激光解吸/电离傅立叶变换离子回旋共振质谱仪(MALALDI-FT-ICRCR-MSI)平台原位分析FFPEPE样品中代谢物含量的方法。该方法包括FFPEPE组织切片，在MALALDI-MSI之前，FFPEPE组织切片经过脱蜡和9-氨基吖啶包衣。利用这一平台，我们以前在FFPEPE样品中检测到了质量范围为m/z 50-1,000的类似1,500 m/z物种；与新鲜冷冻样品相比，重叠部分是m/z物种的72%，表明代谢物在FFPEPE组织样品中大部分是保守的。该方案可以在FFPEPE组织上重复执行，包括组织微阵列和活组织检查等小样本。这一过程可以在一天内完成，这取决于测量的样本大小和使用的栅格大小。这种方法的优点包括样品处理容易、重现性好、高通量和能够在原位显示分子的空间分布。通过该方案获得的数据可用于研究和临床实践。
Formalin-fixed and paraffin-embedded (FFPEPE) tissue specimens are the gold standard for histological examination, and they provide valuable molecular information in tissue-based research. Metabolite assessment from archived tissue samples has not been extensively conducted because of a lack of appropriate protocols and concerns about changes in metabolite content or chemical state due to tissue processing. We present a protocol for the in situ analysis of metabolite content from FFPEPE samples using a high-mass-resolution matrix-assisted laser desorption/ionization fourier-transform ion cyclotron resonance mass spectrometry imaging (MALALDI-FT-ICRCR-MSI) platform. The method involves FFPEPE tissue sections that undergo deparaffinization and matrix coating by 9-aminoacridine before MALALDI-MSI. Using this platform, we previously detected similar to 1,500 m/z species in the mass range m/z 50-1,000 in FFPEPE samples; the overlap compared with fresh frozen samples is 72% of m/z species, indicating that metabolites are largely conserved in FFPEPE tissue samples. This protocol can be reproducibly performed on FFPEPE tissues, including small samples such as tissue microarrays and biopsies. The procedure can be completed in a day, depending on the size of the sample measured and raster size used. Advantages of this approach include easy sample handling, reproducibility, high throughput and the ability to demonstrate molecular spatial distributions in situ. The data acquired with this protocol can be used in research and clinical practice.