A non-instrument-based method for the analysis of formalin-fixed paraffin-embedded human spinal cord via matrix-assisted laser desorption/ionisation imaging mass spectrometry

O'Rourke, MB; Djordjevic, SP; Padula, MP

A non-instrument-based method for the analysis of formalin-fixed paraffin-embedded human spinal cord via matrix-assisted laser desorption/ionisation imaging mass spectrometry

O'Rourke, MB

Djordjevic, SP

Padula, MP

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Publication Type:: Journal Article
Citation:: Rapid Communications in Mass Spectrometry, 2015, 29 (19), pp. 1836 - 1840
Issue Date:: 2015-10-15

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Field	Value	Language
dc.contributor.author	O'Rourke, MB https://orcid.org/0000-0002-0552-2962	en_US
dc.contributor.author	Djordjevic, SP https://orcid.org/0000-0001-9301-5372	en_US
dc.contributor.author	Padula, MP https://orcid.org/0000-0002-8283-0643	en_US
dc.date.available	2015-07-16	en_US
dc.date.issued	2015-10-15	en_US
dc.identifier.citation	Rapid Communications in Mass Spectrometry, 2015, 29 (19), pp. 1836 - 1840	en_US
dc.identifier.issn	0951-4198	en_US
dc.identifier.uri	http://hdl.handle.net/10453/37788
dc.description.abstract	Copyright © 2015 John Wiley & Sons, Ltd. Rationale This paper, in conjunction with a work published earlier this year by O'Rourke et al., aims to provide a comprehensive set of protocols for the analysis of formalin-fixed paraffin-embedded (FFPE) tissue via matrix-assisted laser desorption/ionisation imaging mass spectrometry (MALDI IMS) in a low-cost and highly repeatable and robust way, thereby allowing other research teams to begin their own IMS-centered avenues of research. Methods Samples of FFPE tissue were sectioned at 5 μm, water float mounted to specially prepared ITO glass slides and then dilipidated in a graded alcohol series. Tissue sections were then antigen retrieved under pressure in 20 mmol Tris-HCl (pH 8.8), coated with trypsin and digested O/N at 37°C. Samples were then sublimated with matrix to a final coverage of 0.2 mg/cm<sup>2</sup>, recrystallised at 37 C with 50:50 acetonitrile (ACN)/0.1% trifluoroacetic acid (TFA) for 1 h and analysed with a MALDI TOF/TOF mass spectrometer. Results Serial sections were imaged, revealing little to no variation with regards to image quality and corresponding spectra. We have also attempted to describe the processes that govern the various aspects of this protocol with respect to each step necessary to ensure reproducibility. Conclusions We are confident that this protocol in conjunction with the work published earlier by O'Rourke et al. provides the basis for a repeatable and robust protocol for the analysis of tissues from various sources via MALDI-IMS. The descriptions of key steps within allows for easy adoption of the protocol while allowing for desired modifications to be performed with minimal yet intuitive adjustment.	en_US
dc.relation.ispartof	Rapid Communications in Mass Spectrometry	en_US
dc.relation.isbasedon	10.1002/rcm.7283	en_US
dc.subject.classification	Analytical Chemistry	en_US
dc.subject.mesh	Spinal Cord	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Formaldehyde	en_US
dc.subject.mesh	Trypsin	en_US
dc.subject.mesh	Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization	en_US
dc.subject.mesh	Histocytochemistry	en_US
dc.subject.mesh	Paraffin Embedding	en_US
dc.title	A non-instrument-based method for the analysis of formalin-fixed paraffin-embedded human spinal cord via matrix-assisted laser desorption/ionisation imaging mass spectrometry	en_US
dc.type	Journal Article
utslib.citation.volume	19	en_US
utslib.citation.volume	29	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	03 Chemical Sciences	en_US
utslib.for	04 Earth Sciences	en_US
utslib.for	06 Biological Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	19	en_US
pubs.publication-status	Published	en_US
pubs.volume	29	en_US

Abstract:

Copyright © 2015 John Wiley & Sons, Ltd. Rationale This paper, in conjunction with a work published earlier this year by O'Rourke et al., aims to provide a comprehensive set of protocols for the analysis of formalin-fixed paraffin-embedded (FFPE) tissue via matrix-assisted laser desorption/ionisation imaging mass spectrometry (MALDI IMS) in a low-cost and highly repeatable and robust way, thereby allowing other research teams to begin their own IMS-centered avenues of research. Methods Samples of FFPE tissue were sectioned at 5 μm, water float mounted to specially prepared ITO glass slides and then dilipidated in a graded alcohol series. Tissue sections were then antigen retrieved under pressure in 20 mmol Tris-HCl (pH 8.8), coated with trypsin and digested O/N at 37°C. Samples were then sublimated with matrix to a final coverage of 0.2 mg/cm², recrystallised at 37 C with 50:50 acetonitrile (ACN)/0.1% trifluoroacetic acid (TFA) for 1 h and analysed with a MALDI TOF/TOF mass spectrometer. Results Serial sections were imaged, revealing little to no variation with regards to image quality and corresponding spectra. We have also attempted to describe the processes that govern the various aspects of this protocol with respect to each step necessary to ensure reproducibility. Conclusions We are confident that this protocol in conjunction with the work published earlier by O'Rourke et al. provides the basis for a repeatable and robust protocol for the analysis of tissues from various sources via MALDI-IMS. The descriptions of key steps within allows for easy adoption of the protocol while allowing for desired modifications to be performed with minimal yet intuitive adjustment.

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http://hdl.handle.net/10453/37788