Statistical analysis of time resolved single molecule fluorescence data without time binning

Hinze, G.; Basché, Th.

doi:10.1063/1.3303634

Cited by 14 publications

(11 citation statements)

References 31 publications

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“…More regularly, such measurements have been used to identify temporal heterogeneity, as is natural given the observed FL fluctuations. Minimal binning and bin-free techniques have been applied to assess possible changes in FL over the shortest accessible timescales (69,74,115,122,125,126). Such minimal binning is critical in observing fluctuations in the FL over the course of a given SM trajectory.…”

Section: General Approach and Assessing Heterogeneitymentioning

confidence: 99%

Heterogeneity in Single-Molecule Observables in the Study of Supercooled Liquids

Kaufman

2013

Annu. Rev. Phys. Chem.

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Bulk approaches to studying heterogeneous systems obscure important details, as they report average behavior rather than the distribution of behaviors in such environments. Small-molecule and polymeric supercooled liquids, which display heterogeneity in their dynamics without an underlying structural heterogeneity that sets those dynamics, are important constituents of this category of condensed matter systems. A variety of approaches have been devised to unravel ensemble averaging in supercooled liquids. This review focuses on the ultimate subensemble approach, single-molecule measurements, as they have been applied to the study of supercooled liquids. We detail how three key experimental observables (single-molecule probe rotation, translation, and fluorescence lifetime) have been employed to provide detail on dynamic heterogeneity in supercooled liquids. Special attention is given to the potential for, but also the challenges in, discriminating spatial and temporal heterogeneity and detailing the length scales and timescales of heterogeneity in these systems.

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Section: General Approach and Assessing Heterogeneitymentioning

confidence: 99%

Heterogeneity in Single-Molecule Observables in the Study of Supercooled Liquids

Kaufman

2013

Annu. Rev. Phys. Chem.

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“…However, instead of correlating time binned data, 24 we have employed a statistical analysis without the need of binning the recorded data. 25 Thus the full time resolution of the single photon counts is retained. Note that time binning of the single photon counts either to obtain I d (t) or t(t) would limit the accessible time regime of C d (t) and C f (t) to times t 4 10 À2 s. .…”

Section: Resultsmentioning

confidence: 99%

Single molecule probing of dynamics in supercooled polymers

Hinze

Basché

Vallée

2011

Phys. Chem. Chem. Phys.

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Fluorescence experiments with single BODIPY molecules embedded in a poly(methyl acrylate) matrix have been performed at various temperatures in the supercooled regime. By using pulsed excitation, fluorescence lifetime and linear dichroism time trajectories were accessible at the same time. Both observables have been analyzed without data binning. While the linear dichroism solely reflects single particle dynamics, the fluorescence lifetime observable depends on the molecular environment, so that the dynamics from the polymer host surrounding a chromophore contributes to this quantity. We observe that the lifetime correlation decays slightly faster than polarization correlation, indicating the occurrence of large angular reorientations. Additionally, dichroism time trajectories have been adducted to reveal directly the geometry of rotational dynamics. We identify small but also significantly larger rotational jumps being responsible for the overall molecular reorientation.

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“…By means of a timing signal synchronized to the switching optics, each emitted fluorescence photon can be tagged with the illuminating beam polarization that created it. Such SPC techniques have recently begun to enter molecular biophysics (e.g., in Gopich and Szabo, 2009; Hinze and Basché, 2010; Talaga, 2009; Yang and Xie, 2002). …”

Section: Multiple Channelsmentioning

confidence: 99%

Changepoint Analysis for Single-Molecule Polarized Total Internal Reflection Fluorescence Microscopy Experiments

Beausang

Goldman

2011

Methods in Enzymology

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The experimental study of individual macromolecules has opened a door to determining the details of their mechanochemical operation. Motor enzymes such as the myosin family have been particularly attractive targets for such study, in part because some of them are highly processive and their “product” is spatial motion. But single-molecule resolution comes with its own costs and limitations. Often, the observations rest on single fluorescent dye molecules, which emit a limited number of photons before photobleaching and are subject to complex internal dynamics. Thus, it is important to develop methods that extract the maximum useful information from a finite set of detected photons. We have extended an experimental technique, multiple polarization illumination in total internal reflection fluorescence microscopy (polTIRF), to record the arrival time and polarization state of each individual detected photon. We also extended an analysis technique, previously applied to FRET experiments, that optimally determines times of changes in photon emission rates. Combining these improvements allows us to identify the structural dynamics of a molecular motor (myosin V) with unprecedented detail and temporal resolution.

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Statistical analysis of time resolved single molecule fluorescence data without time binning

Cited by 14 publications

References 31 publications

Heterogeneity in Single-Molecule Observables in the Study of Supercooled Liquids

Heterogeneity in Single-Molecule Observables in the Study of Supercooled Liquids

Single molecule probing of dynamics in supercooled polymers

Changepoint Analysis for Single-Molecule Polarized Total Internal Reflection Fluorescence Microscopy Experiments

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