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  4. Visualization of cell-specific traits in marine picoplankton populations
 
Zitierlink DOI
10.26092/elib/1794

Visualization of cell-specific traits in marine picoplankton populations

Veröffentlichungsdatum
2022-03-14
Autoren
Zeugner, Laura E.  
Betreuer
Fuchs, Bernhard  
Gutachter
Szewzyk, Ulrich  
Zusammenfassung
This thesis investigates the beneficial interplay of advancing technologies in the field of microscopy and microbiology with a focus on the applicability of super-resolution methods for the visualization of sub-cellular structures in prokaryotic cells. DNA, membranes, ribosomal RNA and genes of interest were targeted with specific organic fluorophores or fluorescently-labelled oligo- or polynucleotide probes and visualized on a single-cell level in both, pure cultures and environmental samples. In particular, microbiological techniques based on fluorescence in situ hybridization (FISH) were used, which exploit the hybridization of specific fluorescently-labelled probes to target DNA or ribosomal RNA. The main emphasis was on direct-geneFISH, a tool that was developed to simultaneously detect genes of interest and the 16S rRNA within fixed cells to link microbial identity with a potential function. The image acquisition was performed with four different microscopy methods that, with different approaches, allow for super-resolution: confocal laser scanning microscopy (CLSM) with the Airyscan detector upgrade, structured illumination microscopy (SIM), DNA-point accumulation for imaging in nanoscale topography (PAINT) and stimulated emission depletion (STED) microscopy. Furthermore, this thesis proposes a new approach to quantify individual ribosomes in single cells as a proxy for their growth rate. This is achieved by the visualization of 16S rRNA-derived signals with DNA-PAINT, a method referred to as bacFISH-PAINT.
Schlagwörter
marine microbiology

; 

super-resolution microscopy

; 

fluorescence in situ hybridization

; 

direct-geneFISH
Institution
Universität Bremen  
Fachbereich
Fachbereich 02: Biologie/Chemie (FB 02)  
Dokumenttyp
Dissertation
Lizenz
https://creativecommons.org/licenses/by-sa/4.0/
Sprache
Englisch
Dateien
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PhDthesis_LauraZeugner_PDF-A.pdf

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96.19 MB

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