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Research Article

‘What did I do Wrong?’ an Empirical Evaluation of Sample Preparation Methodologies in Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry Imaging

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Article: FSO362 | Received 02 Oct 2018, Accepted 29 Oct 2018, Published online: 07 Feb 2019
 

Abstract

Aim: This guide aims to broaden the uptake of MALDI-MSI biomedical research by removing the initial ‘lag phase’ associated with empirical determination in sample preparation and data analysis. Methods: Samples from several tissue types were prepared for lipid, protein and peptide MSI analysis. Broadly, samples were cryo sectioned, mounted onto conductive MALDI slides and sublimed with an analyte specific matrix, recrystallised and analyzed in a Bruker UltrafleXtreme MALDI TOF/TOF. Results/conclusion: Here we present a general guide that serves as the first comprehensive, explanatory index for curation and verification of both sample preparation and data generation during the MALDI-MSI process.

Lay abstract

The field of mass spectrometry tissue imaging is a complex field that is designed to provide a map of the molecules on the surface of tissue sections. It often requires a significant investment of time and resources before useful data can be generated; therefore, this paper provides a visual troubleshooting guide that will act as a reference point for a range of sample preparation mistakes and explanations for unusual or suboptimal data. Elimination of the lag phase associated with the development of new techniques will help to expedite the growth and application of this technology.

Supplementary data

To view the supplementary data that accompanythis paper please visit the journal website at: www.tandfonline.com/doi/full/10.4155/fsoa-2018-0095

Acknowledgments

The authors would also like to acknowledge Prof Stuart Cordwell for his expert advice and guidance on the presentation of this manuscript.

Author contributions

MO Rourke collected the majority of data, performed the majority of analysis and wrote the majority of the manuscript. C Smith collected lipid human brain samples and provided analyses and methods for said. B Tse provided thepositive control figure and subsequent analysis. G Sutherland supervised lipid MSI analysis work and incepted brain lipid imaging project and proofread the manuscript. B Crossett provided instrumentation for all lipid imaging and proofread the manuscript. M Padula supervised all imaging performed by MO Rourke, provided instrumentation for said and incepted MSI projects for peptide and protein imaging.

Financial & competing interests disclosure

Tissues were received from the New South Wales Tissue Resource Centre at the University of Sydney and the Sydney Brain Bank, which is supported by the National Health and Medical Research Council of Australia, The University of New South Wales, Neuroscience Research Australia, Schizophrenia Research Institute and the National Institute of Alcohol Abuse and Alcoholism (grant number R24AA012725). The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

Tissues were received from the New South Wales Tissue Resource Centre at the University of Sydney and the Sydney Brain Bank, which is supported by the National Health and Medical Research Council of Australia, The University of New South Wales, Neuroscience Research Australia, Schizophrenia Research Institute and the National Institute of Alcohol Abuse and Alcoholism (grant number R24AA012725). The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.