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  Correlative 3D anatomy and spatial chemistry in animal-microbe symbioses: Developing sample preparation for phase-contrast synchrotron radiation based micro-computed tomography and mass spectrometry imaging

Geier, B., Franke, M., Ruthensteiner, B., González Porras, M., Gruhl, A., Wörmer, L., et al. (2019). Correlative 3D anatomy and spatial chemistry in animal-microbe symbioses: Developing sample preparation for phase-contrast synchrotron radiation based micro-computed tomography and mass spectrometry imaging. Bellingham, Washington: SPIE.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000B-99E1-A Version Permalink: https://hdl.handle.net/21.11116/0000-000B-99E4-7
Genre: Conference Paper

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 Creators:
Geier, B, Author
Franke, M, Author
Ruthensteiner, B, Author
González Porras, MA1, Author                 
Gruhl, A, Author
Wörmer, L, Author
Moosmann, J, Author
Hammel, JU, Author
Dubilier, N, Author
Leisch, N, Author
Liebeke, M, Author
Affiliations:
1External Organizations, ou_persistent22              

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 Abstract: The three-dimensional (3D) architecture of organs, tissues and cells together with the spatial distribution of specific molecules, both enables and drives the close interactions between hosts and microbes. To image the 3D anatomy and molecular composition of an animal-microbe system at a single cell scale we developed a correlative workflow combining phase-contrast synchrotron radiation based micro-computed tomography (PC-SRμCT) and mass spectrometry imaging (MSI). The key challenge in combining both techniques was the synchronization of sample preparation and imaging conditions. We used PC-SRμCT for high-resolution 3D imaging, which is the most suitable x-ray tomographic technique to image soft tissue samples without contrast enhancing agents at a single-cell level. For method development, we used small invertebrates that live in symbiosis with bacteria. For PC-SRμCT samples were paraformaldehyde-fixed and submersed in a buffer-filled capillary. We tested sample preparation for Matrix-assisted laser desorption ionization (MALDI)-MSI after PC-SRμCT and achieved the best results for PFA-fixed samples, processed under cryogenic conditions. The resulting molecular composition permitted imaging of hundreds of different molecule distributions from single tissue sections on a micrometer resolution. Image analysis revealed organ- and cell-specific metabolite distributions, which we could match to corresponding structures in the anatomical 3D PC-SRμCT model.
Our PC-SRμCT-MSI sample preparation- and imaging workflow results in a detailed 3D scenario, which provides an atlas to guide other microscopy or omics approaches towards specific organs and cells. Particularly, for animal-microbe systems this approach presents a powerful tool to visualize anatomical and chemical processes at a microscale, linking structure and function in the symbiosis.

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Language(s): eng - English
 Dates: 2019-09
 Publication Status: Issued
 Pages: 19
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.1117/12.2530652
 Degree: -

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Title: SPIE Optics + Photonics 2019
Place of Event: San Diego, CA, USA
Start-/End Date: 2019-08-11 - 2019-08-15

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Title: Proceedings of SPIE
Source Genre: Series
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Publ. Info: Bellingham, Washington : SPIE
Pages: - Volume / Issue: 11113 Sequence Number: - Start / End Page: - Identifier: ISSN: 0277-786X
CoNE: https://pure.mpg.de/cone/journals/resource/0277-786X

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Title: Developments in X-Ray Tomography XII
Source Genre: Proceedings
 Creator(s):
Müller, B, Editor
Wang, G, Editor
Affiliations:
-
Publ. Info: Bellingham, WA, USA : SPIE
Pages: 19 Volume / Issue: - Sequence Number: - Start / End Page: - Identifier: ISBN: 978-1-5106-2919-6