Localizing and tracking of single molecules with a MINFLUX-microscope for various applications

Weihs, Tobias

dc.contributor.advisor	Hell, Stefan Prof. Dr.
dc.contributor.author	Weihs, Tobias
dc.date.accessioned	2022-01-25T13:51:54Z
dc.date.available	2022-12-08T00:50:10Z
dc.date.issued	2022-01-25
dc.identifier.uri	http://hdl.handle.net/21.11130/00-1735-0000-0008-5A03-0
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-9060
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	530	de
dc.title	Localizing and tracking of single molecules with a MINFLUX-microscope for various applications	de
dc.type	doctoralThesis	de
dc.contributor.referee	Hell, Stefan Prof. Dr.
dc.date.examination	2021-12-10
dc.subject.gok	Physik (PPN621336750)	de
dc.description.abstracteng	Recently MINFLUX microscopy was first published, a technique that localizes single ﬂuorescent molecules by probing with a structured illumination excitation beam featuring a central intensity minimum. Single nanometer precision can be achieved by adaptation of the scan pattern geometry rather than solely relying on an increased number of acquired photons. The technique was transferred from highly specialized optical setups to a general purpose MINFLUX microscope featuring a common ﬂuorescence microscope body. 2D imaging at 1 nm precision and 3D imaging in cells at 2.2 and 1.6 nm lateral and axial precision, respectively, was accomplished, attaining formerly published results. The imaging performance of the microscope was investigated. Live background correction facilitates imaging in cells in higher background situations. For the frst time, 3D MINFLUX tracking was experimentally demonstrated. Sinusoidal bead trajectories induced by a piezo stage and single ﬂuorescent molecules diﬀusing in supported lipid bilayers were tracked in 3D at nanometer precision with 2.8 and 2.1 kHz time resolution, respectively. Super-fast 2D tracking of single ﬂuorescent molecules diﬀusing for several seconds over a micrometer-range was performed. Tracks featuring localization precision of <13 nm at 4.5 kHz and <24 nm with up to 14 kHz time resolution were recorded using only 30 and 9 photons per localization, establishing a new spatio-temporal tracking benchmark for ﬂuorescence microscopy. An analysis framework was developed to extract diﬀusion coeffcients from recorded tracking data. Simulations were employed to investigate the tracking performance of the microscope over a variety of diﬀusion coeffcients using diﬀerent measurement parameters. Diﬀusion coeffcients above 1 µm²/s are within trackable range. Improving the scanner performance of the microscope will shift this limit to higher values.	de
dc.contributor.coReferee	Salditt, Tim Prof. Dr.
dc.contributor.thirdReferee	Köster, Sarah Prof. Dr.
dc.contributor.thirdReferee	Jakobs, Stefan Prof. Dr.
dc.contributor.thirdReferee	Eggeling, Christian Prof. Dr.
dc.contributor.thirdReferee	Egner, Alexander Prof. Dr.
dc.subject.eng	Microscopy	de
dc.subject.eng	Superresolution	de
dc.subject.eng	Fluorescence	de
dc.subject.eng	MINFLUX	de
dc.subject.eng	Tracking	de
dc.identifier.urn	urn:nbn:de:gbv:7-21.11130/00-1735-0000-0008-5A03-0-3
dc.affiliation.institute	Fakultät für Physik	de
dc.description.embargoed	2022-12-08
dc.identifier.ppn	1787154319

Dateien

Name:PhD_thesis_TobiasWeihs_submission.pdf

Größe:107.4Mb

Format:PDF

Beschreibung:PhD thesis

Öffnen

Name:: PhD_thesis_TobiasWeihs_submiss ...
Größe:: 107.4Mb
Format:: PDF
Beschreibung:: PhD thesis

Öffnen

Das Dokument erscheint in:

Fakultät für Physik (inkl. GAUSS) [928]

Zur Kurzanzeige