Molecular morphology of the chick heart visualized by MALDI imaging mass spectrometry.
Molecular morphology of the chick heart visualized by MALDI imaging mass spectrometry.
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DOI：
10.1002/ar.21103
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发表时间：
2010-05
影响因子：
2
通讯作者：
Schey, Kevin L.
中科院分区：
文献类型：
--
作者：
Grey, Angus C.;Gelasco, Andrew K.;Section, Jarren;Moreno-Rodriguez, Ricardo A.;Krug, Edward L.;Schey, Kevin L.
利用MALDI-MS（基质辅助激光解吸/电离质谱法）进行组织成像是一种相对较新的蛋白质组学技术，可同时直接在单个冷冻组织切片内绘制多种蛋白质的空间分布。在这里，我们报告了一种方法的发展，应用MALDI组织成像鸡心脏组织切片固定和石蜡包埋的样品。该方案产生的分子图像可以与高质量的组织学组织切片相关。将灌流的足月鸡心脏固定在酸性乙醇中并包埋在石蜡中。将组织切片（15 μm）收集到导电载玻片上，用二甲苯脱蜡，并转移到水中，用梯度乙醇洗涤，然后风干。在单独的实验中，通过玻璃雾化器的重复循环将三种不同的MALDI基质应用于鸡心脏组织切片。然后使用75-100 μm的光栅步长通过MALDI质谱法分析组织切片，并重建特定m/z比的分子图像。MALDI组织成像揭示了单个心脏切片内的空间分辨蛋白质信号，这些信号对心脏的结构或区域特异，例如，血管、瓣膜、内膜、心肌或隔膜。此外，不需要像免疫组织化学和原位杂交方法那样预先了解蛋白质表达。在单个组织切片内同时定位大量独特蛋白质信号的能力，以及良好的组织学特征保存，为心血管研究人员提供了一种新的工具，以深入了解正常和病理条件下的分子机制。
Utilization of MALDI-MS (matrix-assisted laser desorption/ionization mass spectrometry) for tissue imaging is a relatively new proteomic technique that simultaneously maps the spatial distribution of multiple proteins directly within a single frozen tissue section. Here we report the development of a methodology to apply MALDI tissue imaging to chick heart tissue sections acquired from fixed and paraffin-embedded samples. This protocol produces molecular images that can be related to the high quality histological tissue sections. Perfused term chick hearts were fixed in acidic ethanol and embedded in paraffin wax. Tissue sections (15 μm) were collected onto conductive slides, deparaffinized with xylene and transitioned into water with graded ethanol washes and allowed to air dry. In separate experiments three different MALDI matrices were applied to chick heart tissue sections through repeated cycles from a glass nebulizer. Tissue sections were then analyzed by MALDI mass spectrometry using a raster step-size of 75–100 μm, and molecular images for specific m/z ratios reconstituted. MALDI tissue imaging revealed spatially-resolved protein signals within single heart sections that are specific to structures or regions of the heart, e.g., vessels, valves, endocardium, myocardium, or septa. Moreover, no prior knowledge of protein expression is required as is the case for immunohistochemistry and in situ hybridization methodologies. The ability to simultaneously localize a large number of unique protein signals within a single tissue section, with good preservation of histological features, provides cardiovascular researchers a new tool to give insight into the molecular mechanisms underlying normal and pathological conditions.
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影响因子：
7.4
作者：
Baluya, Dodge L.;Garrett, Timothy J.;Yost, Richard A.
通讯作者：
Yost, Richard A.
影响因子：
48
作者：
Andersson, Malin;Groseclose, M. Reid;Caprioli, Richard M.
通讯作者：
Caprioli, Richard M.
DOI：
10.1016/j.jasms.2005.06.006
发表时间：
2005-10-01
影响因子：
3.2
作者：
Jurchen, JC;Rubakhin, SS;Sweedler, JV
通讯作者：
Sweedler, JV
影响因子：
4.4
作者：
Chaurand, Pierre;Latham, Joey C.;Caprioli, Richard M.
通讯作者：
Caprioli, Richard M.
影响因子：
7.4
作者：
Khatib-Shahidi, Sheerin;Andersson, Malin;Caprioli, Richard M.
通讯作者：
Caprioli, Richard M.