Correlation spectroscopy of individual molecules immobilized on surfaces under ambient conditions

Tietz, C.; Daum, R.; Dräbenstedt, Alexander; Schuster, Jörg; Fleury, L.; Gruber, Achim D.; Wrachtrup, Joerg; Borczyskowski, C. von

doi:10.1016/s0009-2614(97)01243-8

Cited by 9 publications

(11 citation statements)

References 16 publications

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“…A slight deviation of the correlation function from an exponential shape is observed, see Figure 5, similar to the results for sulforhodamine (Tietz et al, 1998). In our measurement procedure, data are acquired from several molecules at different positions in the film, leading to approximately exponential correlation functions, supporting a random character of the fluctuations.…”

Section: Discussionsupporting

confidence: 83%

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Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films

Casoli

Schönhoff

2001

Biological Chemistry

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Fluorescence correlation experiments were performed on rhodamine 6G in PDMS spin-coated films on glass surfaces. With polarised excitation, ensemble bleaching of the dye and single molecule intensity fluctuations were observed. From the statistics of single molecule intensity data taken at different positions in the film, correlation functions were calculated. Two modes of motion with exponential decay shapes and correlation times of tau(c) = 0.15 s and tau(c) = 0.7 s could be detected. Potential origins of intensity fluctuations are lateral diffusion, rotational diffusion or intramolecular fluctuations of dyes involving spectral diffusion or photoinduced processes. From the experimental results, lateral diffusion can be ruled out as a motional mode. Single molecule fluctuations are assigned to changes of the molecular configuration of the dyes, which are rigidly bound to the glass. To assess the environmental influence on such molecular motions, the bulk viscosity of the PDMS was varied over two orders of magnitude, leading to changes of tau(c) of the slow mode by a factor of four. This result proves the sensitivity of the single molecule fluctuations to the molecular scale dynamics of the surrounding polymer matrix and makes the correlation time a measure of the local environment of dye probes.

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Section: Discussionsupporting

confidence: 83%

“…Generally, two distinct timescales of fluctuations could be identified (Lu and Xie, 1997;Tietz et al 1998;Talley and Dunn, 1999). It seems thus likely that the fluctuations observed for Rh6G in PDMS arise from similar effects.…”

Section: Discussionmentioning

confidence: 93%

Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films

Casoli

Schönhoff

2001

Biological Chemistry

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“…24,25 Therefore, we believe that SMS studies of multichromophoric dendrimer systems make it possible to investigate fundamental aspects of energy-transfer processes that are relevant for a number of multichromophoric systems, including photosynthetic complexes and conjugated polymers. 21,22,[26][27][28][29][30][31] Recently, a new type of dendrimer consisting of hexaphenylbenzene building blocks with strongly twisted benzene units was described. [32][33][34] The important feature of the easily soluble 3D polyphenylene dendrimers is the relatively high shape persistence.…”

Section: Introductionmentioning

confidence: 99%

Probing Photophysical Processes in Individual Multichromophoric Dendrimers by Single-Molecule Spectroscopy

et al. 2000

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Individual multichromophoric dendrimer molecules, bearing eight perylenecarboximide chromophores at the rim, immobilized in a thin polyvinylbutyral (PVB) film were studied by far-field fluorescence microscopy. Fluorescence intensity trajectories as a function of time (transients), spectra, and decay traces were recorded separately or simultaneously. For comparison, similar measurements have been performed on a model compound containing one perylenecarboximide chromophore. Collective on/off jumps of the fluorescence intensity were observed for single dendrimer molecules, resembling previously reported collective jumps for the emission of single light-harvesting antenna systems. Spectra and decays of both non-interacting and dimer-like interacting chromophoric sites could be distinguished within an individual dendrimer. Transitions between the different spectral forms and decay times, observed for a single molecule, underline the dynamic character of the interactions among the chromophores. Evidence for a stepwise bleaching process of the multichromophoric system was found. Furthermore, the single-molecule data incontestably prove the assumptions stated in the ensemble model.

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“…This progress has created great enthusiasm in using single fluorescing molecules as sensitive local probes, e.g., in microbiology (11). However, although different features of single-molecule emission such as spontaneous polarization changes (12) and excitation saturation (13) can be interpreted in a straightforward manner, other features such as spontaneous intensity fluctuations on a time scale from milliseconds to seconds or shifts of their fluorescence spectra are not clear (14)(15)(16)(17)(18). A better understanding of these phenomena is required to distinguish between effects associated with the observation of a dye molecule in an intense laser focus and those induced by the interaction of the probe molecule with the local environment.…”

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confidence: 99%

Dynamics of single dye molecules observed by confocal imaging and spectroscopy

1999

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The fluorescence emission of single rhodamine dye molecules (rhodamine 6G and rhodamine 630) at room temperature was analyzed by using scanning confocal laser microscopy in conjunction with polarization analysis, fluorescence spectroscopy, time‐resolved detection (minutes to microseconds), and excitation saturation. Results are presented and discussed 1) for samples with dye molecules at the glass‐air interface and 2) covered with an additional thin protective polymer film (polyvinylbutyral). Under the polymer layer, the single‐molecule fluorescence was more stable than the glass–air interface. This result may be explained by fewer spontaneous variations of the fluorescence rate, polarization changes, spectral shifts, and longer photochemical lifetimes. Cytometry 36:217–223, 1999. © 1999 Wiley‐Liss, Inc.

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Correlation spectroscopy of individual molecules immobilized on surfaces under ambient conditions

Cited by 9 publications

References 16 publications

Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films

Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films

Probing Photophysical Processes in Individual Multichromophoric Dendrimers by Single-Molecule Spectroscopy

Dynamics of single dye molecules observed by confocal imaging and spectroscopy

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