Novel fluorophores for single-molecule imaging

Willets, Katherine A.; Ostroverkhova, Oksana; Hess, Stephan; He, Meng; Twieg, Robert J.; Moerner, W. E.

doi:10.1117/12.506519

Cited by 17 publications

(22 citation statements)

References 22 publications

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“…26, 27 The photobleaching QYs for the Pn-R-F8 molecules embedded in the crystalline t-bu BTBTB host were more than an order of magnitude lower than those in PMMA, depending on the derivative, in part due to a reduced oxygen diffusion in the t-bu BTBTB crystalline environment. High fluorescence QYs and high photostability exhibited by ADT-R-R' and Pn-R-F8 derivatives under study enabled their imaging in a variety of hosts (PMMA, PVK, PS, and t-bu BTBTB) at the single molecule level under wide-field excitation at either 532 nm (ADT-R-R') or 633 nm (Pn-R-F8) (Fig.2b).…”

Section: Resultsmentioning

confidence: 99%

Photophysics of organic semiconductors: from ensemble to the single-molecule level

et al. 2015

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We present photophysical properties of functionalized anthradithiophene (ADT) and pentacene (Pn) derivatives, as well as charge and energy transfer properties of donor-acceptor (D/A) pairs of these derivatives. All molecules studied were fluorescent and photostable enough to be imaged on the single-molecule level in a variety of polymeric and in a functionalized benzothiophene (BTBTB) crystalline host using room-temperature widefield epifluorescence microscopy. Flexibility of functionalization of both guest (ADT, Pn) and host (BTBTB or polymer) molecules can be used for systematic studies of nanoscale morphology and photophysics of D/A organic semiconductor bulk heterojunctions, as well as in applications relying on FRET, using single-molecule fluorescence microscopy.

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Section: Resultsmentioning

confidence: 99%

Photophysics of organic semiconductors: from ensemble to the single-molecule level

et al. 2015

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“…Stimulated by the near order-of-magnitude improvement in quantum yield in the PMMA film compared to toluene solution, a relationship between fluorescence quantum yield and local viscosity or environmental rigidity was proposed. 25 Figure 5 provides two demonstrations of this relationship. In Figure 5A, DCDHF-6 was added to solutions of glycerol/methanol to determine the effect of viscosity on the bulk fluorescence.…”

Section: Environmental Sensitivitymentioning

confidence: 97%

“…In toluene, where the measured quantum yield was quite low, the bulk excited-state lifetime was limited by the response time of the instrument (<400 ps); on the other hand, in PMMA where the quantum yield was near unity, the lifetime was several nanoseconds. 39,41 Bulk studies of DCDHF-6 in different methacrylate polymer hosts revealed that the average excited-state lifetime of the molecule increased with increasing glass transition temperature (T g ) of the host. However, these studies do not directly yield insight into the overall heterogeneity of the environment, since only the average value of the lifetime is measured.…”

Section: Potential Applicationsmentioning

confidence: 99%

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Nonlinear Optical Chromophores as Nanoscale Emitters for Single-Molecule Spectroscopy

et al. 2005

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Fluorescence imaging of single molecules at room temperature is a powerful technique for studying complex condensed phase systems and revealing structure and dynamics hidden by ensemble measurements. Successful single-molecule spectroscopic experiments rely upon strong emitters that can be detected at the level of individual copies above the relevant background signals. This Account discusses a class of nonlinear optical chromophores that not only are well-suited for single-molecule imaging but also offer additional beneficial properties such as a significant ground-state dipole moment, moderate hyperpolarizability, and sensitivity to local environment. An overview of the photophysical properties of several members of this class of molecules as well as a mechanism to help understand the environmental sensitivity is presented. Some preliminary applications of the chromophores as single-molecule reporters in cellular and polymer systems are discussed, along with detection of the emitters by two-photon fluorescence.

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“…Total internal reflection fluorescence ͑TIRF͒ 20 resolved some of these problems and allowed the application to myosin II and V in vitro. [21][22][23][24][25][26] It is possible to further decrease axial dimension and avoid reflections and scattering by the use of a surface plasmon assisted microscope ͑SPAM͒.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of a single cross-bridge in familial hypertrophic cardiomyopathy heart muscle measured by reverse Kretschmann fluorescence

et al. 2010

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Abstract. Familial hypertrophic cardiomyopathy ͑FHC͒ is a serious heart disease that often leads to a sudden cardiac death of young athletes. It is believed that the alteration of the kinetics of interaction between actin and myosin causes FHC by making the heart to pump blood inefficiently. We set out to check this hypothesis ex vivo. During contraction of heart muscle, a myosin cross-bridge imparts periodic force impulses to actin. The impulses are analyzed by fluorescence correlation spectroscopy ͑FCS͒ of fluorescently labeled actin. To minimize observation volume and background fluorescence, we carry out FCS measurements in surface plasmon coupled emission mode in a reverse Kretschmann configuration. Fluorescence is a result of near-field coupling of fluorophores excited in the vicinity of the metal-coated surface of a coverslip with the surface plasmons propagating in the metal. Surface plasmons decouple on opposite sides of the metal film and emit in a directional manner as far-field p-polarized radiation. We show that the rate of changes of orientation is significantly faster in contracting cardiac myofibrils of transgenic mice than wild type. These results are consistent with the fact that mutated heart muscle myosin translates actin faster in in vitro motility assays. © 2010Society of Photo-Optical Instrumentation Engineers.

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Novel fluorophores for single-molecule imaging

Cited by 17 publications

References 22 publications

Photophysics of organic semiconductors: from ensemble to the single-molecule level

Photophysics of organic semiconductors: from ensemble to the single-molecule level

Nonlinear Optical Chromophores as Nanoscale Emitters for Single-Molecule Spectroscopy

Kinetics of a single cross-bridge in familial hypertrophic cardiomyopathy heart muscle measured by reverse Kretschmann fluorescence

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