Yang Junhai, Caprioli Richard M
Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA.
J Mass Spectrom. 2014 May;49(5):417-22. doi: 10.1002/jms.3354.
We have developed matrix pre-coated targets for imaging proteins in thin tissue sections by matrix-assisted laser desorption/ionization mass spectrometry. Gold covered microscope slides were coated with sinapinic acid (SA) in batches in advance and were shown to be stable for over 6 months when kept in the dark. The sample preparation protocol using these SA pre-coated targets involves treatment with diisopropylethylamine (DIEA)-H2 O vapor, transforming the matrix layer to a viscous ionic liquid. This SA-DIEA ionic liquid layer extracts proteins and other analytes from tissue sections that are thaw mounted to this target. DIEA is removed by the immersion of the target into diluted acetic acid, allowing SA to co-crystallize with extracted analytes directly on the target. Ion images (3-70 kDa) of sections of mouse brain and rat kidney at spatial resolution down to 10 µm were obtained. Use of pre-coated slides greatly reduces sample preparation time for matrix-assisted laser desorption/ionization imaging while providing high throughput, low cost and high spatial resolution images.
我们已经开发出用于通过基质辅助激光解吸/电离质谱对薄组织切片中的蛋白质进行成像的基质预涂覆靶材。预先批量在金覆盖的显微镜载玻片上涂覆芥子酸(SA),并且在黑暗中保存时显示出6个月以上的稳定性。使用这些SA预涂覆靶材的样品制备方案包括用二异丙基乙胺(DIEA)-H2O蒸汽处理,将基质层转化为粘性离子液体。这种SA-DIEA离子液体层从解冻后安装到该靶材上的组织切片中提取蛋白质和其他分析物。通过将靶材浸入稀释的乙酸中来去除DIEA,使SA与提取的分析物直接在靶材上共结晶。获得了小鼠脑和大鼠肾切片的离子图像(3-70 kDa),空间分辨率低至10 µm。使用预涂覆载玻片大大减少了基质辅助激光解吸/电离成像的样品制备时间,同时提供高通量、低成本和高空间分辨率的图像。