Optimal Preparation of Formalin Fixed Samples for Peptide Based Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Workflows

被引：7

作者：

O'Rourke, Matthew B. ^{[1
,2
]}

Padula, Matthew P. ^{[2
]}

Smith, Caine ^{[3
]}

Youssef, Priscilla ^{[4
]}

Cordwell, Stuart ^{[1
]}

Witting, Paul ^{[4
]}

Sutherland, Greg ^{[3
]}

Crossett, Ben ^{[1
]}

机构：

[1] Univ Sydney, Mass Spectrometry Core Facil, Sydney, NSW, Australia

[2] Univ Technol Sydney, Prote Core Facil, Sydney, NSW, Australia

[3] Univ Sydney, Sch Med Sci, Discipline Pathol, Neuropathol Grp, Sydney, NSW, Australia

[4] Univ Sydney, Sch Med Sci, Discipline Pathol, Redox Biol Grp, Sydney, NSW, Australia

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 131期

关键词：

Biochemistry; Issue; 131; Mass spectrometry Imaging; matrix assisted laser desorption ionization; formalin fixed paraffin embedded; tissue Imaging; MALDI MSI; sublimation; TISSUE; PROTEINS;

D O I：

10.3791/56778

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

The use of matrix-assisted laser desorption/ionization, mass spectrometry imaging (MALDI MSI) has rapidly expanded, since this technique analyzes a host of biomolecules from drugs and lipids to N-glycans. Although various sample preparation techniques exist, detecting peptides from formaldehyde preserved tissues remains one of the most difficult challenges for this type of mass spectrometric analysis. For this reason, we have created and optimized a robust methodology that preserves the spatial information contained within the sample, while eliciting the greatest number of ionizable peptides. We have also aimed to achieve this in a cost effective and simple way, thereby eliminating potential bias or preparation error, which can occur when using automated instrumentation. The end result is a reproducible and inexpensive protocol.

引用

页数：7

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