Improving Tissue Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging. Part 1: Using Microspotting
Improving Tissue Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging. Part 1: Using Microspotting
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DOI：
10.1021/ac901328p
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发表时间：
2009-10-01
影响因子：
7.4
通讯作者：
Fournier, Isabelle
中科院分区：
文献类型：
--
作者：
Franck, Julien;Arafah, Karim;Fournier, Isabelle
基质辅助激光解吸电离质谱成像（MALDI MSI）是目前获得组织切片中内源性和外源性分子分布的一种有效技术。因此，它可以用于研究分子在不同生理阶段的进化，以找到标记或获得信号通路的知识。为了提供有价值的信息，我们必须仔细控制样品制备，以避免在此步骤中组织内感兴趣的分子的任何离域。目前，可以使用两种策略来将化学品（例如MALDI基质）存款到组织上，这两种策略都涉及将掉落到表面上的微滴的产生。第一种策略涉及溶液的微喷雾。在这里，我们一直感兴趣的微点战略的发展，其中纳米液滴的溶剂被喷射的压电装置在组织水平上产生微点。这样的系统允许人们通过创建斑点阵列来精确地控制样品制备。在基质结晶方面，微量点样MALDI基质与经典（移液）方法的结果几乎不相容。因此，我们合成和研究了新的固体离子基质，以获得高的分析性能，使用这样的沉积系统。由于在这些条件下产生的印刷的高稳定性，这些发展使得能够优化准备时间。我们还研究了用于进行组织上消化的微点样，以便鉴定蛋白质或从福尔马林固定和石蜡包埋（FFPE）组织样品中进行工作。我们已经表明，微点是一个有趣的方法对组织消化。在这种特定的制备策略下研究了肽、蛋白质和脂质，以改善这类分子的成像性能。
Nowadays, matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) is a powerful technique to obtain the distribution of endogenous and exogenous molecules within tissue sections. It can, thus, be used to study the evolution of molecules across different physiological stages in order to find out markers or get knowledge on signaling pathways. In order to provide valuable information, we must carefully control the sample preparation to avoid any delocalization of molecules of interest inside the tissue during this step. Currently, two strategies can be used to deposit chemicals, such as the MALDI matrix, onto the tissue both involving generation of microdroplets that will be dropped off onto the surface. First strategy involves microspraying of solutions. Here, we have been interested in the development of a microspotting strategy, where nanodroplets of solvent are ejected by a piezoelectric device to generate microspots at the tissue level. Such systems allow one to precisely control sample preparation by creating an array of spots. In terms of matrix crystallization, a microspotting MALDI matrix is hardly compatible with the results by classical (pipetting) methods. We have thus synthesized and studied new solid ionic matrixes in order to obtain high analytical performance using such a deposition system. These developments have enabled optimization of the preparation time because of the high stability of the printing that is generated in these conditions. We have also studied microspotting for performing on-tissue digestion in order to go for identification of proteins or to work from formalin fixed and paraffin embedded (FFPE) tissue samples. We have shown that microspotting is an interesting approach for on tissue digestion. Peptides, proteins, and lipids were studied under this specific preparation strategy to improve imaging performances for this class of molecules.