Fluorescence fluctuation studies of biomolecular interactions in solutions, biomembranes and live cells
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Fluorescence spectroscopy and imaging have a very broad spectrum of applicationswithin the life sciences, in particular for detection and characterization ofbiomolecular dynamics and interactions in different environments. This thesis comprisesprojects that strive to further expand the information content extracted fromthe detected fluorescence, leading to sensitive readout parameters for studies ofbiomolecular dynamics and interactions. Two major strategies are presented toachieve this aim. The first strategy is based on the expansion of the availablereadout parameters beyond the "traditional" fluorescence parameters: intensity,wavelength, polarization and fluorescence lifetime. The additional parameters arebased on blinking properties of fluorescent labels. In particular on transitions betweensinglet and triplet states, and transitions between the trans- and cis-isomersof fluorophores. Two publications in the thesis are based on this strategy (paperI and IV). The second strategy is based on the utilization of fluorescence intensityfluctuations in order to detect the oligomerization mechanisms of fluorescentlylabeled peptides and proteins. This strategy combines the intensity fluctuationanalysis and the readout of distance dependent energy transfer between fluorescentmolecules together with the correlation analysis of fluorescence from two labeledproteins emitting at different wavelengths. Another two publications presented inthe thesis are based on the second comprehensive strategy (papers II and III).The work presented in this thesis shows that the blinking kinetics of fluorescentlabels contain significant information that can be exploited by a combination of fluctuationsanalysis with distance dependent excitation energy transfer between thefluorescent molecules, or by analysis of fluorescence covariance between moleculesthat emit at different wavelengths. These fluorescence-based methods have a significantpotential for molecular interaction studies in the biomedical field.
Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2016. , x, 59 p.
TRITA-FYS, ISSN 0280-316X ; 2016:22
FCS, FCCS, Isomerization, TRAST, NMR, FRET, biomombrane, fluidity
Research subject Biological Physics
IdentifiersURN: urn:nbn:se:kth:diva-187708ISBN: 978-91-7729-026-1OAI: oai:DiVA.org:kth-187708DiVA: diva2:931216
2016-06-13, FB52, KTH, AlbaNova University Center, Roslagsvägen 30 B, Stockholm, 13:00 (English)
Delon, Antoine, Professor
Widengren, Jerker, Professor
QC 201605272016-05-272016-05-262016-05-31Bibliographically approved
List of papers