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dc.contributor.author
Kunz, Leo
dc.contributor.supervisor
Schroeder, Timm
dc.contributor.supervisor
Škoda, Radek
dc.contributor.supervisor
Reddy, Sai
dc.date.accessioned
2020-09-25T13:43:48Z
dc.date.available
2019-09-24T09:41:34Z
dc.date.available
2019-09-24T10:39:41Z
dc.date.available
2020-09-25T13:43:48Z
dc.date.issued
2019
dc.identifier.uri
http://hdl.handle.net/20.500.11850/366084
dc.identifier.doi
10.3929/ethz-b-000366084
dc.description.abstract
Multicolour 3D quantitative imaging of large tissue volumes is pivotal to understand the development and organization of tissues as well as interactions of cells and the distribution of secreted molecules in situ. However, in situ imaging is technically challenging. In particular, the preparation and imaging of large bone and marrow sections, as well as faithful imaging and quantification of individual secreted molecules is not straightforward or possible with current techniques. The localization of most molecules in situ therefore remains unknown. In the work presented in this thesis, I developed together with my colleagues an integrated pipeline to generate reproducible high-dimensional quantitative data from bone marrow and any other tissue. Further, I present my efforts in adapting the Proximity Ligation Assay for large volume in situ imaging of individual molecules, including new 3D quantification strategies and software. Using these approaches, we measured the spatial relationship between hematopoietic cells, bone surfaces and Schwann cells within bone marrow. We further created an atlas of nonhematopoietic cells in the bone marrow and I investigated the process of metastatic colonialization of the bone marrow. I then looked into the distribution of individual C-X-C motif ligand 12 (CXCL12) molecules and found broad CXCL12 distribution with local enrichment at bone surfaces, but no long-range intra-marrow CXCL12 gradients, correcting current assumptions about migration control of hematopoietic stem cells in bone marrow. Overall the here presented technique developments and adaptations will enable new discoveries down to the molecular level in situ. While the biological findings answer some of the current questions in bone marrow biology, the questions raised by my findings represent the basis for more research in this new branch of quantitative molecular in situ research.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
ETH Zurich
en_US
dc.rights.uri
http://rightsstatements.org/page/InC-NC/1.0/
dc.subject
Imaging
en_US
dc.subject
Immunostaining
en_US
dc.subject
Cytokine
en_US
dc.subject
quantitative image analysis
en_US
dc.subject
BONE MARROW (CYTOLOGY, HISTOLOGY)
en_US
dc.subject
Blood
en_US
dc.title
Development and application of quantitative in situ imaging techniques
en_US
dc.type
Doctoral Thesis
dc.rights.license
In Copyright - Non-Commercial Use Permitted
dc.date.published
2019-09-24
ethz.size
209 p.
en_US
ethz.code.ddc
DDC - DDC::6 - Technology, medicine and applied sciences::610 - Medical sciences, medicine
en_US
ethz.identifier.diss
26188
en_US
ethz.publication.place
Zurich
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02060 - Dep. Biosysteme / Dep. of Biosystems Science and Eng.::03992 - Schroeder, Timm / Schroeder, Timm
en_US
ethz.date.deposited
2019-09-24T09:41:47Z
ethz.source
FORM
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.date.embargoend
2020-09-24
ethz.rosetta.installDate
2019-09-24T10:47:23Z
ethz.rosetta.lastUpdated
2019-09-24T10:47:23Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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