Navigation überspringen
Universitätsbibliothek Heidelberg
Status: Bibliographieeintrag

Verfügbarkeit
Standort: ---
Exemplare: ---
heiBIB
 Online-Ressource
Verfasst von:Liesche, Clarissa [VerfasserIn]   i
 Grußmayer, Kristin Stefanie [VerfasserIn]   i
 Ludwig, Michael [VerfasserIn]   i
 Wörz, Stefan [VerfasserIn]   i
 Rohr, Karl [VerfasserIn]   i
 Herten, Dirk-Peter [VerfasserIn]   i
 Beaudouin, Joël [VerfasserIn]   i
 Eils, Roland [VerfasserIn]   i
Titel:Automated analysis of single-molecule photobleaching data by statistical modeling of spot populations
Verf.angabe:Clarissa Liesche, Kristin S. Grußmayer, Michael Ludwig, Stefan Wörz, Karl Rohr, Dirk-Peter Herten, Joël Beaudouin, and Roland Eils
E-Jahr:2015
Jahr:1 December 2015
Umfang:11 S.
Fussnoten:Gesehen am 23.06.2020
Titel Quelle:Enthalten in: Biophysical journal
Ort Quelle:Cambridge, Mass. : Cell Press, 1960
Jahr Quelle:2015
Band/Heft Quelle:109(2015), 11, Seite 2352-2362
ISSN Quelle:1542-0086
Abstract:The number of fluorophores within a molecule complex can be revealed by single-molecule photobleaching imaging. A widely applied strategy to analyze intensity traces over time is the quantification of photobleaching step counts. However, several factors can limit and bias the detection of photobleaching steps, including noise, high numbers of fluorophores, and the possibility that several photobleaching events occur almost simultaneously. In this study, we propose a new approach, to our knowledge, to determine the fluorophore number that correlates the intensity decay of a population of molecule complexes with the decay of the number of visible complexes. We validated our approach using single and fourfold Atto-labeled DNA strands. As an example we estimated the subunit stoichiometry of soluble CD95L using GFP fusion proteins. To assess the precision of our method we performed in silico experiments showing that the estimates are not biased for experimentally observed intensity fluctuations and that the relative precision remains constant with increasing number of fluorophores. In case of fractional fluorescent labeling, our simulations predicted that the fluorophore number estimate corresponds to the product of the true fluorophore number with the labeling fraction. Our method, denoted by spot number and intensity correlation (SONIC), is fully automated, robust to noise, and does not require the counting of photobleaching events.
DOI:doi:10.1016/j.bpj.2015.10.035
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext: https://doi.org/10.1016/j.bpj.2015.10.035
 Volltext: http://www.sciencedirect.com/science/article/pii/S000634951501111X
 DOI: https://doi.org/10.1016/j.bpj.2015.10.035
Datenträger:Online-Ressource
Sprache:eng
K10plus-PPN:1701940019
Verknüpfungen:→ Zeitschrift

Permanenter Link auf diesen Titel (bookmarkfähig):  https://katalog.ub.uni-heidelberg.de/titel/68590013   QR-Code
zum Seitenanfang