Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz

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Nina Höller

Nariae Baik-Schneditz

Bernhard Schwaberger

Lukas Peter Mileder

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Földes-Papp, Z.
"True" single-molecule molecule observations by fluorescence correlation spectroscopy and two-color fluorescence cross-correlation spectroscopy.
Exp Mol Pathol. 2007; 82(2):147-155 Doi: 10.1016/j.yexmp.2006.12.002
Web of Science PubMed FullText FullText_MUG

Führende Autor*innen der Med Uni Graz: Földes-Papp Zeno
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Abstract:: Fluorescence correlation spectroscopy (FCS) and two-color fluorescence cross-correlation spectroscopy (FCCS) are a measure of fluctuations of detected light as a fluorescence molecule diffuses through a femtoliter detection volume caused by a tightly focused laser and confocal optics. Fluorescence from a single molecule can easily be distinguished from the slight background associated with a femtoliter of solvent. At a solution concentration of about 1 nM, the probability that there is an analyte molecule in the probe volume is less than one. Although fluorescence from individual molecules is collected, the data are analyzed by autocorrelation or two-color cross-correlation functions that are the average of thousands of molecules. Properties of single molecules are not obtained. I have been working on problems and opportunities associated with very dilute solutions. The molecule in the confocal probe volume is most probably the molecule that just diffused out, turned around, and diffused back in, i.e., reentered. For the first time, some theoretical results of the novel theory of the meaningful time are presented that enable study of just one single molecule over extended periods of times without immobilization or hydrodynamic focusing. Reentries that may also be called reoccurrences or encounters of a single molecule are significant because during measurement times they give rise to fluctuation phenomena such as molecule number fluctuations. Likewise, four criteria have been developed that can be used to verify that there is only one "selfsame" molecule in the laser probe volume during the experiment: (Földes-Papp, Z., 2006. What it means to measure a single molecule in a solution by fluorescence fluctuation spectroscopy. Exp. Mol. Pathol. 80 (3) 209-218).
Find related publications in this database (using NLM MeSH Indexing): Color -; Diffusion -; Fluorescent Dyes -; Models, Biological -; Solutions -; Spectrometry, Fluorescence - methods
Find related publications in this database (Keywords): single molecule; solution; individual molecule; selfsame molecule; selfsame single-molecule regime; selfsame molecule likelihood estimator; meaningful time theory of single-molecule analysis; fluorescence fluctuation microscopy; fluorescence correlation spectroscopy (FCS); two-color fluorescence cross-correlation spectroscopy (FCCS); single (solution)-phase single-molecule auto; and two-color cross-correlation spectroscopy (SPSM-FCS)