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Method Article
Label-free Imaging of Biomolecules in Murine Brain Sections Using the 3D OrbiSIMS
Ian Gilmore
National Physical Laboratory, , NiCE-MSI, Teddington, Middlesex, UK.
Corresponding Author
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A step-by-step protocol for label-free imaging of biomolecules, including lipids and neurotransmitters, in murine brain sections using the 3D OrbiSIMS is provided. The protocol describes requirements for mounting of murine brain sections, the calibration of the Orbitrap™ mass spectrometer, the set-up of the imaging acquisition, and the analysis of the hyperspectral imaging dataset to generate biomolecule distribution maps. Following the protocol, the distribution of biomolecules with molecular weights in the range from 80 to 1200 Da can be imaged at the microscale with spatial resolution < 2 μm (< 1.5 μm can be achieved) , high mass resolving power (m/Δm) > 200,000 at m/z 200 and mass accuracy < 2 ppm. The protocol may be completed within 5 hours. This protocol accompanies Passarelli et al (Nature Methods, 10.1038/nmeth.4504, published online 13 November 2017).
Keywords
Biomedical imaging, label free, molecular pathology, secondary ion mass spectrometry, Orbitrap™, OrbiSIMS
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Table 1 Table 1: Positive and negative secondary ions used for Orbitrap MS calibration.
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Table 2 Minimum time taken to acquire an image in minutes depending on mass resolving power and image resolution.
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Associated Publications
The 3D OrbiSIMS—label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power
by Melissa K Passarelli, Alexander Pirkl, Rudolf Moellers, +13
Nature Methods (04 December, 2017)
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This is a preprint. It has not completed peer review.
Method Article
Label-free Imaging of Biomolecules in Murine Brain Sections Using the 3D OrbiSIMS
Ian Gilmore
National Physical Laboratory, , NiCE-MSI, Teddington, Middlesex, UK.
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 04 Dec, 2017
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Biological techniques
License:
This work is licensed under a CC BY 4.0 License. Read Full License
A step-by-step protocol for label-free imaging of biomolecules, including lipids and neurotransmitters, in murine brain sections using the 3D OrbiSIMS is provided. The protocol describes requirements for mounting of murine brain sections, the calibration of the Orbitrap™ mass spectrometer, the set-up of the imaging acquisition, and the analysis of the hyperspectral imaging dataset to generate biomolecule distribution maps. Following the protocol, the distribution of biomolecules with molecular weights in the range from 80 to 1200 Da can be imaged at the microscale with spatial resolution < 2 μm (< 1.5 μm can be achieved) , high mass resolving power (m/Δm) > 200,000 at m/z 200 and mass accuracy < 2 ppm. The protocol may be completed within 5 hours. This protocol accompanies Passarelli et al (Nature Methods, 10.1038/nmeth.4504, published online 13 November 2017).
Associated Publications
The 3D OrbiSIMS—label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power
by Melissa K Passarelli, Alexander Pirkl, Rudolf Moellers, +13
Nature Methods (04 December, 2017)