Time-dependent evolution of tissue markers by MALDI-MS imaging
Time-dependent evolution of tissue markers by MALDI-MS imaging
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DOI：
10.1002/pmic.200800201
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发表时间：
2008-09-01
期刊：
影响因子：
3.4
通讯作者：
Pitt, Andrew R.
中科院分区：
文献类型：
--
作者：
Goodwin, Richard J. A.;Dungworth, Jessica C.;Pitt, Andrew R.
我们已经使用MALDI-MS成像（MALDI-MSI）来监测当组织切片被快速带到室温短到中等时间段时的信号的时间依赖性外观和损失。在低温恒温切片机中切割小鼠脑切片，置于MALDI靶上并使其升温至室温30秒至3小时。然后将切片重新冷冻，通过乙醇处理固定并通过MALDI-MSI进行分析。观察到一系列标志物的强度在整个时间过程中变化，既增加又减少，其中一些标志物的强度在30秒内显著变化，并且标志物还显示出组织位置特异性演变。将这种自溶产生的标志物与在可比的16小时组织上胰蛋白酶消化中进化的标志物直接进行比较，并且观察到在两项研究中进化的标志物是显著不同的。这些变化提供了重要的额外水平的位置依赖性信息，用于映射变化和寻找组织中的疾病依赖性生物标志物。它们还表明，需要相当小心，以允许比较MALDI-MSI实验之间的生物标志物，并且还对将多个组织切片解冻安装到MALDI-MS靶标上的标准实践具有影响。
We have used MALDI-MS imaging (MALDI-MSI) to monitor the time dependent appearance and loss of signals when tissue slices are brought rapidly to room temperature for short to medium periods of time. Sections from mouse brain were cut in a cryostat microtome, placed on-a MALDI target and allowed to warm to room temperature for 30 s to 3 h. Sections were then refrozen, fixed by ethanol treatment and analysed by MALDI-MSI. The intensity of a range of markers were seen to vary across the time course, both increasing and decreasing, with the intensity of some markers changing significantly within 30 s and markers also showed tissue location specific evolution. The markers resulting from this autolysis were compared directly to those that evolved in a comparable 16 h on-tissue trypsin digest, and the markers that evolved in the two studies were seen to be substantially different. These changes offer an important additional level of location-dependent information for mapping changes and seeking disease-dependent biomarkers in the tissue. They also indicate that considerable care is required to allow comparison of biomarkers between MALDI-MSI experiments and also has implications for the standard practice of thaw-mounting multiple tissue sections onto MALDI-MS targets.