The molecular scanner in microscope mode
The molecular scanner in microscope mode
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DOI：
10.1002/rcm.2747
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发表时间：
2006-01-01
影响因子：
2
通讯作者：
Heeren, Ron M. A.
中科院分区：
文献类型：
--
作者：
Luxembourg, Stefan L.;Vaezaddeh, Ali R.;Heeren, Ron M. A.
探讨了显微镜模式基质辅助激光解吸/电离（MALDI）成像质谱（IMS）与蛋白质鉴定方法——分子扫描仪的结合。分子扫描方法提高了在显微镜模式下检测和鉴定高质量蛋白质的灵敏度。采用十二烷基硫酸钠/聚丙烯酰胺凝胶电泳（SDS-PAGE）对分离后的蛋白质分布进行了验证。在聚偏二氟乙烯捕获膜上涂覆MALDI样品后，在TRIFT-II离子显微镜上记录了高质量、高空间分辨率的离子图像。该组合方法的灵敏度估计为5 pmol。鉴定所需的最小样品消耗量估计大于100 fmol。开发了软件工具来分析光谱数据，并生成宽质量范围和单分子组分显微镜模式离子图像和单质量电荷比微探针模式图像。版权所有(c) 2006约翰威利父子有限公司
The combination of microscope mode matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) with protein identification methodology: the molecular scanner, was explored. The molecular scanner approach provides improvement of sensitivity of detection and identification of high-mass proteins in microscope mode IMS. The methodology was tested on protein distributions obtained after separation by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). High-quality, high-spatial-resolution ion images were recorded on a TRIFT-II ion microscope after gold coating of the MALDI sample preparation on the poly(vinylidenedifluoride) capture membranes. The sensitivity of the combined method is estimated to be 5 pmol. The minimum amount of sample consumed, needed for identification, was estimated to be better than 100 fmol. Software tools were developed to analyze the spectral data and to generate broad mass range and single molecular component microscope mode ion images and single mass-to-charge ratio microprobe mode images. Copyright (c) 2006 John Wiley & Sons, Ltd.