Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †

James W. Hughes; Emma K. Sisley; Oliver J. Hale; Helen J. Cooper

doi:10.1039/d3sc04933g

Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †

James W. Hughes, Emma K. Sisley, Oliver J. Hale, Helen J. Cooper^*

^*Corresponding author for this work

Biosciences

Research output: Contribution to journal › Article › peer-review

Abstract

Previously, we have shown that native ambient mass spectrometry imaging allows the spatial mapping of folded proteins and their complexes in thin tissue sections. Subsequent top-down native ambient mass spectrometry of adjacent tissue section enables protein identification. The challenges associated with protein identification by this approach are (i) the low abundance of proteins in tissue and associated long data acquisition timescales and (ii) irregular spatial distributions which hamper targeted sampling of the relevant tissue location. Here, we demonstrate that these challenges may be overcome through integration of laser capture microdissection in the workflow. We show identification of intact protein assemblies in rat liver tissue and apply the approach to identification of proteins in the granular layer of rat cerebellum.

Original language	English
Journal	Chemical Science
Early online date	7 Mar 2024
DOIs	https://doi.org/10.1039/d3sc04933g
Publication status	E-pub ahead of print - 7 Mar 2024

Bibliographical note

Acknowledgments:
H. J. C is an EPSRC Established Career Fellow (EP/S002979/1). J. W. H and O. J. H were funded by EPSRC (EP/S002979/1), and E. K. S was funded by the University of Birmingham. The Orbitrap Eclipse mass spectrometer used in this work was funded by BBSRC (BB/S019456/1). The authors thank Prof. Richard Goodwin (AstraZeneca) for providing the tissue samples, Dr Alessandro di Maio and Ms Rebecca Gascoyne (Microscopy and Imaging Services and Birmingham University) for help and training on the Zeiss PALM MicroBeam, and Jen Kulvinder (Institute for Biomedical Research) for help with H&E staining.

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10.1039/d3sc04933gLicence: Creative Commons: Attribution (CC BY)

A new mass spectrometer for structural proteomics and protein imaging
Thomas, C. (Co-Investigator), Grant, M. (Co-Investigator), Lovering, A. (Co-Investigator), Alderwick, L. (Co-Investigator), Tomlinson, M. (Co-Investigator), Winn, P. (Co-Investigator), Knowles, T. (Co-Investigator), Cooper, H. (Principal Investigator), Styles, I. (Co-Investigator), Gibbs, D. (Co-Investigator), Huber, D. (Co-Investigator), Bhatt, A. (Co-Investigator) & Leney, A. (Co-Investigator)
Biotechnology & Biological Sciences Research Council
1/07/19 → 30/06/20
Project: Research
NAMS: Native ambient mass spectrometry
Cooper, H. (Principal Investigator)
Engineering & Physical Science Research Council
1/06/19 → 30/11/23
Project: Research

Cite this

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title = "Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †",

abstract = "Previously, we have shown that native ambient mass spectrometry imaging allows the spatial mapping of folded proteins and their complexes in thin tissue sections. Subsequent top-down native ambient mass spectrometry of adjacent tissue section enables protein identification. The challenges associated with protein identification by this approach are (i) the low abundance of proteins in tissue and associated long data acquisition timescales and (ii) irregular spatial distributions which hamper targeted sampling of the relevant tissue location. Here, we demonstrate that these challenges may be overcome through integration of laser capture microdissection in the workflow. We show identification of intact protein assemblies in rat liver tissue and apply the approach to identification of proteins in the granular layer of rat cerebellum.",

author = "James W. Hughes and Sisley, {Emma K.} and Hale, {Oliver J.} and Cooper, {Helen J.}",

note = "Acknowledgments: H. J. C is an EPSRC Established Career Fellow (EP/S002979/1). J. W. H and O. J. H were funded by EPSRC (EP/S002979/1), and E. K. S was funded by the University of Birmingham. The Orbitrap Eclipse mass spectrometer used in this work was funded by BBSRC (BB/S019456/1). The authors thank Prof. Richard Goodwin (AstraZeneca) for providing the tissue samples, Dr Alessandro di Maio and Ms Rebecca Gascoyne (Microscopy and Imaging Services and Birmingham University) for help and training on the Zeiss PALM MicroBeam, and Jen Kulvinder (Institute for Biomedical Research) for help with H&E staining.",

year = "2024",

month = mar,

day = "7",

doi = "10.1039/d3sc04933g",

language = "English",

journal = "Chemical Science",

issn = "2041-6520",

publisher = "Royal Society of Chemistry",

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T1 - Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †

AU - Hughes, James W.

AU - Sisley, Emma K.

AU - Hale, Oliver J.

AU - Cooper, Helen J.

N1 - Acknowledgments: H. J. C is an EPSRC Established Career Fellow (EP/S002979/1). J. W. H and O. J. H were funded by EPSRC (EP/S002979/1), and E. K. S was funded by the University of Birmingham. The Orbitrap Eclipse mass spectrometer used in this work was funded by BBSRC (BB/S019456/1). The authors thank Prof. Richard Goodwin (AstraZeneca) for providing the tissue samples, Dr Alessandro di Maio and Ms Rebecca Gascoyne (Microscopy and Imaging Services and Birmingham University) for help and training on the Zeiss PALM MicroBeam, and Jen Kulvinder (Institute for Biomedical Research) for help with H&E staining.

PY - 2024/3/7

Y1 - 2024/3/7

N2 - Previously, we have shown that native ambient mass spectrometry imaging allows the spatial mapping of folded proteins and their complexes in thin tissue sections. Subsequent top-down native ambient mass spectrometry of adjacent tissue section enables protein identification. The challenges associated with protein identification by this approach are (i) the low abundance of proteins in tissue and associated long data acquisition timescales and (ii) irregular spatial distributions which hamper targeted sampling of the relevant tissue location. Here, we demonstrate that these challenges may be overcome through integration of laser capture microdissection in the workflow. We show identification of intact protein assemblies in rat liver tissue and apply the approach to identification of proteins in the granular layer of rat cerebellum.

AB - Previously, we have shown that native ambient mass spectrometry imaging allows the spatial mapping of folded proteins and their complexes in thin tissue sections. Subsequent top-down native ambient mass spectrometry of adjacent tissue section enables protein identification. The challenges associated with protein identification by this approach are (i) the low abundance of proteins in tissue and associated long data acquisition timescales and (ii) irregular spatial distributions which hamper targeted sampling of the relevant tissue location. Here, we demonstrate that these challenges may be overcome through integration of laser capture microdissection in the workflow. We show identification of intact protein assemblies in rat liver tissue and apply the approach to identification of proteins in the granular layer of rat cerebellum.

U2 - 10.1039/d3sc04933g

DO - 10.1039/d3sc04933g

M3 - Article

SN - 2041-6520

JO - Chemical Science

JF - Chemical Science

ER -

Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †

Abstract

Bibliographical note

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Projects

A new mass spectrometer for structural proteomics and protein imaging

NAMS: Native ambient mass spectrometry

Cite this