Luxembourg Stefan L, Vaezaddeh Ali R, Amstalden Erika R, Zimmermann-Ivol Catherine G, Hochstrasser Denis F, Heeren Ron M A
FOM Institute for Atomic and Molecular Physics, Kruislaan 407, 1098 SJ Amsterdam, The Netherlands.
Rapid Commun Mass Spectrom. 2006;20(22):3435-42. doi: 10.1002/rcm.2747.
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.
探索了显微镜模式下的基质辅助激光解吸/电离（MALDI）成像质谱（IMS）与蛋白质鉴定方法（分子扫描仪）的结合。分子扫描仪方法提高了显微镜模式IMS中高质量蛋白质检测和鉴定的灵敏度。该方法在通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳（SDS-PAGE）分离后获得的蛋白质分布上进行了测试。在聚偏二氟乙烯捕获膜上进行MALDI样品制备并镀上金后，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皮摩尔。鉴定所需消耗的最小样品量估计优于100飞摩尔。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成分的显微镜模式离子图像以及单质荷比微探针模式图像。
