Variable-angle ultraviolet total-internal-reflection fluorescence spectroscopy using a white excitation light source

Toriumi, Mitsuhiro; Saito, S.; Kawaguchi, Kakuhiro; Aiki, K.

doi:10.1063/1.1145464

Cited by 7 publications

(5 citation statements)

References 24 publications

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“…Where detailed spectroscopic information is required from TIRF, commercial spectrouorometers are readily adapted. For most applications one of the wide range of commercially available TIRF spectrouorometer cells would be appropriate -however the literature provides examples of custom-built apparatus 47 including for specialised applications such as high pressures. 48 The main developmental thrust in TIRF imaging in recent years has been to improve the lateral resolution below the 'diffraction limit' (which is typically around 300 nm).…”

Section: Emission Spectroscopymentioning

confidence: 99%

Total internal reflection spectroscopy for studying soft matter

Woods

Bain

2014

Soft Matter

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Total internal reflection (TIR) spectroscopy is a widely used technique to study soft matter at interfaces. This tutorial review aims to provide researchers with an overview of the principles, experimental design and applications of TIR spectroscopy to enable them to understand how this class of techniques might be used in their research. It also highlights limitations and pitfalls of TIR techniques, which will assist readers in critically analysing the literature. Techniques covered include attenuated total reflection infrared spectroscopy (ATR-IR), TIR fluorescence, TIR Raman scattering and cavity-enhanced techniques. Other related techniques are briefly described.

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Section: Emission Spectroscopymentioning

confidence: 99%

Total internal reflection spectroscopy for studying soft matter

Woods

Bain

2014

Soft Matter

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“…VA-TIRF has been used to map out depth-dependent concentration profiles of fluorescein and fluorescein-labeled IgG next to surfaces [85], pyrene in thin polymer films [96], and fluorescein and acridine orange next to surfaces [97]. This method has also been used to map out the topology of cell-substrate contact regions by applying the technology to adherent cells with fluorescently labeled cytoplasms or membranes [89,94,[98][99][100][101][102][103][104].…”

Section: Variable Incidence Anglesmentioning

confidence: 99%

Total Internal Reflection Fluorescence Microscopy: Applications in Biophysics

Thompson

Pero

2005

Springer Series on Fluorescence

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“…Other authors took advantage of the symmetry of a cylindrical prism [2,12,13] or used a spherical lens (J. P. Henry, personal communication). VA-EW imaging has been used to study static fluorophore distributions, dye adsorption at interfaces [14][15][16] or cell-substrate separation distances [11,[17][18][19]. For studies of dynamic processes, e.g.…”

Section: A Simple Optical Configuration For Reproducible Beam Scanningmentioning

confidence: 99%

REVIEW Imaging Transmitter Release. II. A Practical Guide to Evanescent-wave Imaging

Oheim

2001

Lasers Med Sci

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Total internal reflection of a laser beam at an interface between two media of different refractive index sets up an evanescent wave field in the medium with lower refractive index. This near field decays over a distance of approximately lambda/5, lambda denoting the wavelength of light, and thus provides a convenient means for the confinement of fluorescence excitation to the near-interface region. Evanescent-wave excitation thereby permits, for example, the observation of individual fluorophores at the surface despite the presence of high concentrations in bulk solution. Although evanescent-wave excitation of fluorescence and the related technique of surface-plasmon resonance have a long record in the study of chemical reactions at surfaces, adsorption kinetics or spectroscopy, their potential for biomedical studies is only gradually emerging. Evanescent-wave microscopy provides high-contrast images of the near-membrane region of cells grown on a glass substrate at unprecedented resolution. At present, no commercial equipment is available for evanescent-wave microscopy. This review aims at readers who want to modify their fluorescence microscope to include an evanescent-wave illumination mode. Starting from the point that every objective exceeding a certain numerical aperture is generating evanescent waves, we demonstrate how the optical path can be modified to suppress the far-field excitation, and how one can switch easily between these types of illumination. The phenomena resulting from interactions of evanescent waves with cells are reviewed. The ways in which systematic variations of the angle of incident light can be used to obtain quantitative information on fluorophore distance, distribution and concentration are also discussed.

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Variable-angle ultraviolet total-internal-reflection fluorescence spectroscopy using a white excitation light source

Cited by 7 publications

References 24 publications

Total internal reflection spectroscopy for studying soft matter

Total internal reflection spectroscopy for studying soft matter

Total Internal Reflection Fluorescence Microscopy: Applications in Biophysics

REVIEW Imaging Transmitter Release. II. A Practical Guide to Evanescent-wave Imaging

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