Exploring Fluorescence Antibunching in Solution To Determine the Stoichiometry of Molecular Complexes

0089208 - ÚFCH JH 2008 RIV US eng J - Journal Article
Sýkora, Jan - Kaiser, K. - Gregor, I. - Bönigk, W. - Schmalzing, G. - Enderlain, J.
Exploring Fluorescence Antibunching in Solution To Determine the Stoichiometry of Molecular Complexes.
[Měření “antibunchingu“ fluorescence v roztoku: cesta k určení stechiometrie molekulovýxch komplexů.]
Analytical Chemistry. Roč. 79, - (2007), s. 4040-4049. ISSN 0003-2700. E-ISSN 1520-6882
R&D Projects: GA MŠMT(CZ) LC06063
Institutional research plan: CEZ:AV0Z40400503
Keywords : exploring fluorescence antibunching * molecular complexes * biophysical methods
Subject RIV: CF - Physical ; Theoretical Chemistry
Impact factor: 5.287, year: 2007

Fluorescence antibunching is a well-known technique for determining the number of independent emitters per molecule or molecular complex. It was rarely applied to autofluorescent proteins due to the necessity of collecting large numbers of fluorescence photons from a single molecule, which is usually impossible to achieve with rather photolabile autofluorescent proteins. Here, we measure fluorescence antibunching on molecules in solution, allowing us to accumulate data over a large number of molecules. We use that method for determining an average stoichiometry of molecular complexes. The proposed method is absolute in the sense that it does not need any calibration or referencing. We develop the necessary theoretical background and check the method on pure dye solutions and on molecular complexes with known stoichiometry.

Fluorescenční antibunching je známá metoda pro určování počtu nezávislých emiterů přítomných v molekulárních komplexech. Tato metoda byla ale zřídka aplikována pro studium autofluorescenčních proteinů imobilizovaných na povrchu kvůli jejich nízké fotostabilitě. V této práci jsme měřili fluorescenční antibunching v roztoku a nasbírali tak data z obrovského počtu molekul, což nám umožnilo určit stechiometrii komplexů těchto fotolabilních proteinů. Tato metoda navíc nevyžaduje kalibraci standardem.
Permanent Link: http://hdl.handle.net/11104/0150490