Storage Conditions of Human Kidney Tissue Sections Affect Spatial Lipidomics Analysis Reproducibility.
Storage Conditions of Human Kidney Tissue Sections Affect Spatial Lipidomics Analysis Reproducibility.
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DOI：
10.1021/jasms.0c00256
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发表时间：
2020-12-02
影响因子：
3.2
通讯作者：
Kidney Precision Medicine Project
中科院分区：
文献类型：
--
作者：
Lukowski JK;Pamreddy A;Velickovic D;Zhang G;Pasa-Tolic L;Alexandrov T;Sharma K;Anderton CR;Kidney Precision Medicine Project
脂质通常是不稳定的，不稳定的，并且倾向于随着时间的推移而降解，因此研究这些分子在其最天然状态下的最重要性。我们试图了解人肾组织切片的空间脂质组学分析的最佳储存条件。具体而言，我们评估了人类肾脏组织切片在几个不同的日子在整个跨度的一周内使用我们建立的协议阐明脂质使用高质量分辨率基质辅助激光解吸/电离质谱成像（MALDI-MSI）。我们研究了在五种不同条件下储存的肾组织切片：在-80 °C下开放储存，真空密封并在-80 °C下储存，在-80 °C下储存前预涂基质，在氮气气氛下储存在-80 °C下，以及在室温下在干燥器中储存。将结果与从冷冻切片后立即制备和分析的肾组织切片获得的数据进行比较。使用METASPACE处理数据。在储存一周后，储存在室温下的切片显示出最大量的脂质降解，而储存在氮气和-80 °C下的切片保留了与新鲜切割组织相关的最大数量的重叠注释。总体而言，我们发现，无论储存条件如何，组织切片的分子降解随着时间的推移是不可避免的，但是在低温下将组织切片储存在惰性气体中可以减少组织切片内的分子降解。
Lipids often are labile, unstable, and tend to degrade overtime, so it is of the upmost importance to study these molecules in their most native state. We sought to understand the optimal storage conditions for spatial lipidomic analysis of human kidney tissue sections. Specifically, we evaluated human kidney tissue sections on several different days throughout the span of a week using our established protocol for elucidating lipids using high mass resolution matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). We studied kidney tissue sections stored under five different conditions: open stored at −80 °C, vacuumed sealed and stored at −80 °C, with matrix preapplied before storage at −80 °C, under a nitrogen atmosphere and stored at −80 °C, and at room temperature in a desiccator. Results were compared to data obtained from kidney tissue sections that were prepared and analyzed immediately after cryosectioning. Data was processed using METASPACE. After a week of storage, the sections stored at room temperature showed the largest amount of lipid degradation, while sections stored under nitrogen and at −80 °C retained the greatest number of overlapping annotations in relation to freshly cut tissue. Overall, we found that molecular degradation of the tissue sections was unavoidable over time, regardless of storage conditions, but storing tissue sections in an inert gas at low temperatures can curtail molecular degradation within tissue sections.
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通过质谱法对组织中脂质生物化学的MALDI成像。
影响因子：
62.1
作者：
Berry, Karin A. Zemski;Hankin, Joseph A.;Barkley, Robert M.;Spraggins, Jeffrey M.;Caprioli, Richard M.;Murphy, Robert C.
通讯作者：
Murphy, Robert C.
影响因子：
3.4
作者：
Goodwin, Richard J. A.;Dungworth, Jessica C.;Pitt, Andrew R.
通讯作者：
Pitt, Andrew R.
影响因子：
4.3
作者：
Dill, Allison L.;Eberlin, Livia S.;Cooks, R. Graham
通讯作者：
Cooks, R. Graham
影响因子：
3.8
作者：
Holland, NT;Pfleger, L;Bastaki, M
通讯作者：
Bastaki, M
影响因子：
48
作者：
Cornett, Dale S.;Reyzer, Michelle L.;Caprioli, Richard M.
通讯作者：
Caprioli, Richard M.