dc.contributor.advisor | Enderlein, Jörg Prof. Dr. | |
dc.contributor.author | Mojiri, Soheil | |
dc.date.accessioned | 2021-01-07T11:23:38Z | |
dc.date.available | 2021-10-19T00:50:12Z | |
dc.date.issued | 2021-01-07 | |
dc.identifier.uri | http://hdl.handle.net/21.11130/00-1735-0000-0005-153A-3 | |
dc.identifier.uri | http://dx.doi.org/10.53846/goediss-8391 | |
dc.identifier.uri | http://dx.doi.org/10.53846/goediss-8391 | |
dc.language.iso | eng | de |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
dc.subject.ddc | 530 | de |
dc.title | Advances in enhanced multi-plane 3D imaging and image scanning microscopy | de |
dc.type | doctoralThesis | de |
dc.contributor.referee | Enderlein, Jörg Prof. Dr. | |
dc.date.examination | 2020-11-24 | |
dc.subject.gok | Physik (PPN621336750) | de |
dc.description.abstracteng | High-speed 3D optical microscopy is an indispensable requirement
in studying rapid processes like signaling in neuronal networks, flagellar
motion, or complex motion of living and highly dynamic subcellular
components. However, the 3D imaging capability often compromises
either the acquisition speed or the complexity of the imaging
system. Multi-plane imaging offers a parallelized acquisition of
several focal planes at the same time, which tremendously enhances
the temporal resolution. Nonetheless, most of the current multi-plane
approaches are complex and require several optical elements to create
multiple beams with different optical path lengths, limiting their
broad application.
To address these problems, in this thesis, employing a multi-plane
prism, we introduce three novel techniques that each improve the concept
of rapid 3D optical microscopy in a different direction: First, we
accomplish a multi-plane detection for a phase-contrast microscope
resulting in a 3D, label-free, and non-complex imaging system with
a ~ 4ms of temporal resolution. We applied this system in real-time
studying isolated bare axonemes of Chlamydomonas beating in the
vicinity of a surface. This enabled us to observe their non-zero torsional
motion where the torsion sign slowly changes from negative at
the basal end toward positive at the distal end of flagella.
Second, straightforwardly, we combine an experimental spectral unmixing
setup with the multi-plane detection and obtain an instant 3-
color 3D microscope with a minimal axial color aberration of 140 nm
in a ~ 2.5 μm axial range. The performance of such a microscope is
verified in volumetric imaging of three different subcellular components
in fixed COS-7 cells.
Third, a single-color multi-plane fluorescence microscope has been
employed as a rapid 3D particle tracking velocimetry method in microscale
to study single particles tracing the so-called Marangoni flow.
Using that, besides the velocity fields of single particles, we achieve a
localization precision of ~ 60 nm in the lateral and ~ 300 nm in the axial
direction over a 120 120 13 m3 field of view. Altogether, using
these three techniques, we extend the temporal resolution, throughput,
and applicability of 3D optical microscopy.
In another project, we implemented image scanning microscopy simultaneously
in two colors on a conventional wide-field microscope
by applying an LED light source and a digital micro-mirror device.
This permits us to realize an inexpensive, speckle noise-free, and
easy-to-implement super-resolution add-on for a conventional epi-fluorescence
microscope. | de |
dc.contributor.coReferee | Neef, Andreas Dr. | |
dc.contributor.thirdReferee | Wouters, Fred Prof. Dr. | |
dc.contributor.thirdReferee | Egner, Alexander Prof. Dr. | |
dc.contributor.thirdReferee | Stark, Holger Prof. Dr. | |
dc.contributor.thirdReferee | Betz, Timo Prof. Dr. | |
dc.subject.eng | Optical microscopy | de |
dc.subject.eng | 3D imaging | de |
dc.subject.eng | Multi-plane imaging | de |
dc.subject.eng | Phase-contrast microscopy | de |
dc.subject.eng | Chlamydomonas flagella | de |
dc.subject.eng | Spectral unmixing | de |
dc.subject.eng | Micro-scale particle imaging velocimetry | de |
dc.subject.eng | Marangoni flow | de |
dc.subject.eng | Image scanning microscopy | de |
dc.identifier.urn | urn:nbn:de:gbv:7-21.11130/00-1735-0000-0005-153A-3-5 | |
dc.affiliation.institute | Fakultät für Physik | de |
dc.description.embargoed | 2021-10-18 | |
dc.identifier.ppn | 1744211221 | |