{"id":{"repo_id":"heid-diss","oai_identifier":"oai:archiv.ub.uni-heidelberg.de:5033"},"canonical_url":"https://search.dev.ndltd.org/etd/heid-diss/oai:archiv.ub.uni-heidelberg.de:5033","repository":{"repo_id":"heid-diss","name":"Universität Heidelberg","base_url":"http://archiv.ub.uni-heidelberg.de/volltextserver/cgi/oai2"},"display":{"title":"Fluorescence Depletion by Stimulated Emission in Single-Molecule Spectroscopy","abstract":"This thesis presents the first application of the stimulated emission depletion (STED) technique to the field of single-molecule fluorescence spectroscopy. It is demonstrated that fluorescence quenching induced by STED is reversible and can be repeated a large number of cycles on a single molecule. Being ideal point-like probes, single molecules can therefore be used to characterize the resolution of STED microscopes. In a spectroscopic study, two simplifed models of the photophysical processes involved in STED are analyzed and applied to the experimental determination of the stimulated emission cross sections on a single-molecule level. In addition, the STED concept as applied in subdiffraction-resolution microscopy is transferred to fluorescence fluctuation spectroscopy. A successful implementation promises to expand the possibilities particularly of fluorescence correlation spectroscopy (FCS) which is already the most widely used fluctuation technique today but is restricted to concentrations on the nanomolar scale. Reducing the detection volume beyond the diffraction limit could render the micromolar range accessible and could thus open up new applications in the life sciences.","abstract_html":"This thesis presents the first application of the stimulated emission depletion (STED) technique to the field of single-molecule fluorescence spectroscopy. It is demonstrated that fluorescence quenching induced by STED is reversible and can be repeated a large number of cycles on a single molecule. Being ideal point-like probes, single molecules can therefore be used to characterize the resolution of STED microscopes. In a spectroscopic study, two simplifed models of the photophysical processes involved in STED are analyzed and applied to the experimental determination of the stimulated emission cross sections on a single-molecule level. In addition, the STED concept as applied in subdiffraction-resolution microscopy is transferred to fluorescence fluctuation spectroscopy. A successful implementation promises to expand the possibilities particularly of fluorescence correlation spectroscopy (FCS) which is already the most widely used fluctuation technique today but is restricted to concentrations on the nanomolar scale. Reducing the detection volume beyond the diffraction limit could render the micromolar range accessible and could thus open up new applications in the life sciences.","abstract_has_math":false,"creators":["Kastrup, Lars"],"institution":"Universität Heidelberg","degree_name":null,"degree_level":"thesis.doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Wolfrum, Jürgen"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2004,"date_issued":"2004-10-22","date_published":"2004-10-22","updated_at":"2026-07-24T02:29:58Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://www.ub.uni-heidelberg.de/archiv/5033","outbound_label":"Repository record","outbound_source":"source_url"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Wolfrum, Jürgen"]},{"key":"dc:creator","label":"Author","values":["Kastrup, Lars"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:publisher","label":"Institution","values":["Universitätsbibliothek Heidelberg"]},{"key":"dc:type","label":"Dc Type","values":["doctoralThesis"]},{"key":"thesis:degree_level","label":"Degree Level","values":["thesis.doctoral"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Universität Heidelberg"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["This thesis presents the first application of the stimulated emission depletion (STED) technique to the field of single-molecule fluorescence spectroscopy. It is demonstrated that fluorescence quenching induced by STED is reversible and can be repeated a large number of cycles on a single molecule. Being ideal point-like probes, single molecules can therefore be used to characterize the resolution of STED microscopes. In a spectroscopic study, two simplifed models of the photophysical processes involved in STED are analyzed and applied to the experimental determination of the stimulated emission cross sections on a single-molecule level. In addition, the STED concept as applied in subdiffraction-resolution microscopy is transferred to fluorescence fluctuation spectroscopy. A successful implementation promises to expand the possibilities particularly of fluorescence correlation spectroscopy (FCS) which is already the most widely used fluctuation technique today but is restricted to concentrations on the nanomolar scale. Reducing the detection volume beyond the diffraction limit could render the micromolar range accessible and could thus open up new applications in the life sciences.","Diese Arbeit stellt die erste Anwendung der Fluoreszenzlöschung durch stimulierte Emission (stimulated emission depletion, STED) auf das Gebiet der Einzelmolekül-Fluoreszenzspektroskopie dar. Es wird gezeigt, dass STED an einem einzelnen Molekül reversibel ist und eine Vielzahl von Zyklen wiederholt werden kann. Als ideale punktförmige Strahler können einzelne Moleküle daher eingesetzt werden, um das Auflösungsvermögen von STED-Mikroskopen zu charakterisieren. In einer spektroskopischen Studie werden zwei vereinfachte Modelle der an STED beteiligten photophysikalischen Prozesse analysiert and zur experimentellen Bestimmung der Wirkungsquerschnitte für stimulierte Emission auf Einzelmolekülbasis herangezogen. Außerdem wird das STED-Konzept aus der optischen Mikroskopie zur Durchbrechung der Beugungsrenze auf die Fluoreszenzfluktuationsspektroskopie übertragen. Eine erfolgreiche Umsetzung verspricht vor allem, die Möglichkeiten der Fluoreszenzkorrelationsspektroskopie (FCS) zu erweitern, die bereits heute die meistgenutzte Fluktuationsmethode darstellt, jedoch bislang auf nanomolare Konzentrationen beschränkt ist. Die Verkleinerung des Detektionsvolumens auf Größen unterhalb der optischen Beugungsgrenze könnte den mikromolaren Konzentrationsbereich und damit neue Anwendungen im Bereich der Life Sciences erschließen."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Fluorescence Depletion by Stimulated Emission in Single-Molecule Spectroscopy","Fluoreszenzlöschung durch Stimulierte Emission in der Einzelmolekülspektroskopie"]}]}],"canonical_facts":{"dc:contributor":["Wolfrum, Jürgen"],"dc:creator":["Kastrup, Lars"],"dc:description.abstract":["This thesis presents the first application of the stimulated emission depletion (STED) technique to the field of single-molecule fluorescence spectroscopy. It is demonstrated that fluorescence quenching induced by STED is reversible and can be repeated a large number of cycles on a single molecule. Being ideal point-like probes, single molecules can therefore be used to characterize the resolution of STED microscopes. In a spectroscopic study, two simplifed models of the photophysical processes involved in STED are analyzed and applied to the experimental determination of the stimulated emission cross sections on a single-molecule level. In addition, the STED concept as applied in subdiffraction-resolution microscopy is transferred to fluorescence fluctuation spectroscopy. A successful implementation promises to expand the possibilities particularly of fluorescence correlation spectroscopy (FCS) which is already the most widely used fluctuation technique today but is restricted to concentrations on the nanomolar scale. Reducing the detection volume beyond the diffraction limit could render the micromolar range accessible and could thus open up new applications in the life sciences.","Diese Arbeit stellt die erste Anwendung der Fluoreszenzlöschung durch stimulierte Emission (stimulated emission depletion, STED) auf das Gebiet der Einzelmolekül-Fluoreszenzspektroskopie dar. Es wird gezeigt, dass STED an einem einzelnen Molekül reversibel ist und eine Vielzahl von Zyklen wiederholt werden kann. Als ideale punktförmige Strahler können einzelne Moleküle daher eingesetzt werden, um das Auflösungsvermögen von STED-Mikroskopen zu charakterisieren. In einer spektroskopischen Studie werden zwei vereinfachte Modelle der an STED beteiligten photophysikalischen Prozesse analysiert and zur experimentellen Bestimmung der Wirkungsquerschnitte für stimulierte Emission auf Einzelmolekülbasis herangezogen. Außerdem wird das STED-Konzept aus der optischen Mikroskopie zur Durchbrechung der Beugungsrenze auf die Fluoreszenzfluktuationsspektroskopie übertragen. Eine erfolgreiche Umsetzung verspricht vor allem, die Möglichkeiten der Fluoreszenzkorrelationsspektroskopie (FCS) zu erweitern, die bereits heute die meistgenutzte Fluktuationsmethode darstellt, jedoch bislang auf nanomolare Konzentrationen beschränkt ist. Die Verkleinerung des Detektionsvolumens auf Größen unterhalb der optischen Beugungsgrenze könnte den mikromolaren Konzentrationsbereich und damit neue Anwendungen im Bereich der Life Sciences erschließen."],"dc:format.medium":["application/pdf"],"dc:publisher":["Universitätsbibliothek Heidelberg"],"dc:title":["Fluorescence Depletion by Stimulated Emission in Single-Molecule Spectroscopy","Fluoreszenzlöschung durch Stimulierte Emission in der Einzelmolekülspektroskopie"],"dc:type":["doctoralThesis"],"thesis:degree_level":["thesis.doctoral"],"thesis:institution_name":["Universität Heidelberg"]},"updated_at":"2026-07-24T02:29:58Z"}