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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry
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Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry

机译：DAN基质的升华用于大鼠脑组织中神经节苷脂的检测和可视化用于MALDI成像质谱

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摘要

Sample preparation is key for optimal detection and visualization of analytes in Matrix-assisted Laser Desorption/Ionization (MALDI) Imaging Mass Spectrometry (IMS) experiments. Determining the appropriate protocol to follow throughout the sample preparation process can be difficult as each step must be optimized to comply with the unique characteristics of the analytes of interest. This process involves not only finding a compatible matrix that can desorb and ionize the molecules of interest efficiently, but also selecting the appropriate matrix deposition technique. For example, a wet matrix deposition technique, which entails dissolving a matrix in solvent, is superior for desorption of most proteins and peptides, whereas dry matrix deposition techniques are particularly effective for ionization of lipids. Sublimation has been reported as a highly efficient method of dry matrix deposition for the detection of lipids in tissue by MALDI IMS due to the homogeneity of matrix crystal deposition and minimal analyte delocalization as compared to many wet deposition methods ,. Broadly, it involves placing a sample and powdered matrix in a vacuum-sealed chamber with the samples pressed against a cold surface. The apparatus is then lowered into a heated bath (sand or oil), resulting in sublimation of the powdered matrix onto the cooled tissue sample surface. Here we describe a sublimation protocol using 1,5-diaminonaphthalene (DAN) matrix for the detection and visualization of gangliosides in the rat brain using MALDI IMS.
机译：样品制备是在基质辅助激光解吸/电离（MALDI）成像质谱（IMS）实验中最佳检测和可视化分析物的关键。确定适合整个样品制备过程的方案可能很困难，因为必须优化每个步骤以符合目标分析物的独特特征。该过程不仅涉及寻找可以有效解吸和离子化目标分子的相容基质，还包括选择合适的基质沉积技术。例如，需要将基质溶解在溶剂中的湿基质沉积技术对于大多数蛋白质和肽的解吸是优越的，而干基质沉积技术对于脂质的电离特别有效。与许多湿式沉积方法相比，由于基质晶体沉积的均质性和最小的分析物离位，升华已被报道为通过MALDI IMS检测组织中脂质的一种高效的干式基质沉积方法。广义上讲，它涉及将样品和粉末状基质放在真空密封的室内，并将样品压在冷的表面上。然后将设备降低到加热浴（砂或油）中，使粉末状基质升华到冷却的组织样本表面上。在这里，我们描述了一种使用1,5-二氨基萘（DAN）矩阵的升华协议，用于使用MALDI IMS检测和可视化大鼠脑中神经节苷脂。

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