| dc.contributor.advisor | Jakobs, Stefan Prof. Dr. | |
| dc.contributor.author | Schnorrenberg, Sebastian | |
| dc.date.accessioned | 2018-04-23T10:09:05Z | |
| dc.date.available | 2018-04-23T10:09:05Z | |
| dc.date.issued | 2018-04-23 | |
| dc.identifier.uri | http://hdl.handle.net/11858/00-1735-0000-002E-E3C4-2 | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-6838 | |
| dc.language.iso | eng | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject.ddc | 570 | de |
| dc.title | In vivo super-resolution live-cell RESOLFT-microscopy of Drosophila melanogaster and Arabidopsis thaliana | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Jakobs, Stefan Prof. Dr. | |
| dc.date.examination | 2017-08-15 | |
| dc.description.abstracteng | The optical resolution of conventional light microscopy is limited by the diffraction
barrier of light. This resolution limit impedes the characterization of cellular
structures featuring a size smaller than half the wavelength of light. RESOLFT
(reversible saturable optical linear fluorescence transitions)-microscopy breaks the
diffraction barrier by targeted swichting of fluorophores between separable states
using low light levels. Since its first demonstration in 2005, RESOLFT-microscopy
has proved its usability in high resolution imaging of living single cells.
This work reports, for the first time, the establishment of RESOLFT microscopy in
a multicellular organism, namely Drosophila melanogaster and its first application
for in vivo-imaging.
Using RESOLFT-microscopy on isolated tissues, a lateral resolution of 50 - 60 nm
on single microtubule filaments was achieved. Additionally, the dynamic changes of
the microtubule network were captured in high spatial resolution by recording 40
consecutive RESOLFT frames. The fluorescent probe rsEGFP2 enabled high resolution
three dimensional imaging with 4 fold improved axial resolution. Using intact
living Drosophila melanogaster larvae, in vivo high resolution RESOLFT imaging
with a resolution of ~ 50 nm could be demonstrated, which up to now had not been
shown.
Moreover, this work reports on the first application of RESOLFT-microscopy in
the plant Arabidopsis thaliana. The relatively low light levels used in RESOLFTmicroscopy
enabled high resolution imaging in this light absorbing model organism
of plant research. RESOLFT imaging of green leaf epidermis cells resulted in lateral
resolution of ~ 70 nm. Further, it could be shown that the autofluorescence
of chloroplasts can be excluded in the fluorescence recordings of rsEGFP2 using
fluorescence lifetime gating. Time-lapse recording facilitated the characterization of
the dynamic behaviour of microtubule filaments in epidermis cells over a period of
approximately 1 hour without any signs of phototoxicity. | de |
| dc.contributor.coReferee | Shcherbata, Halyna Dr. | |
| dc.subject.eng | high-resolution microscopy | de |
| dc.subject.eng | Drosophila melanogaster | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-11858/00-1735-0000-002E-E3C4-2-5 | |
| dc.affiliation.institute | Biologische Fakultät für Biologie und Psychologie | de |
| dc.subject.gokfull | Biologie (PPN619462639) | de |
| dc.identifier.ppn | 1019392266 | |