In Situ Imaging of Tryptic Peptides by MALDI Imaging Mass Spectrometry Using Fresh-Frozen or Formalin-Fixed, Paraffin-Embedded Tissue.
In Situ Imaging of Tryptic Peptides by MALDI Imaging Mass Spectrometry Using Fresh-Frozen or Formalin-Fixed, Paraffin-Embedded Tissue.
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DOI：
10.1002/cpps.65
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发表时间：
2018-11
影响因子：
--
通讯作者：
Drake RR
中科院分区：
文献类型：
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作者：
Angel PM;Norris-Caneda K;Drake RR
胰多肽成像是基质辅助激光解吸/电离成像质谱仪(MALDI IMS)的主要工作流程，并导致了高度多元化的蛋白质定位的新信息。过去几年的技术进步为基质和酶的自动喷洒提供了强大的工具。当结合高质量分辨率和高质量精度时，现在的仪器研究通常会导致远远超过1,000个多肽峰的二维图谱。本方案描述了酶和基质的样品制备、喷雾和应用，以及从新鲜冰冻(FF)或福尔马林固定的石蜡包埋(FFPE)组织切片中二维绘制胰解肽的MALDI FT-ICR仪器考虑因素。还描述了从组织切片中提取胰蛋白酶多肽以进行LC-MS/MS鉴定的方法。
Tryptic peptide imaging is a primary workflow for matrix-assisted laser desorption/ ionization imaging mass spectrometry (MALDI IMS) and has led to new information reporting highly multiplexed protein localization. Technological advances within the last few years have produced robust tools for automated spraying of both matrix and enzymes. When combined with high mass resolution and high mass accuracy instrumentation studies now generally result in two dimensional mapping of well over 1,000 peptide peaks. This protocol describes sample preparation, spraying and application of enzymes and matrices, and MALDI FT-ICR instrumental considerations for two dimensional mapping of tryptic peptides from fresh frozen (FF) or formalin-fixed, paraffin-embedded (FFPE) tissue sections. Procedures for extraction of tryptic peptides from tissue sections for LC-MS/MS identification are also described.