Total internal reflection fluorescence spectroscopy

D. Horsley, J. Herron, V. Hlady, and J. D. Andrade, Human and hen lysozyme adsorption A comparative study using total internal reflection fluorescence spectroscopy and molecular graphics, in Proteins at Interfaces Physicochemical and Biochemical Studies (J. L. Brash and T. A. Horbett, eds.), ACS Symposium Series No. 343, pp. 290-305, American Chemical Society, Washington, D.C. (1987). 

V. Hlady, C. Golander, and J. D. Andrade, Hydrophobicity gradient on silica surfaces A study using total internal reflection fluorescence spectroscopy, Colloids Surf. 33, 185-190... 

M. Masuhara, S. Tazuke, N. Tamai, and I. Yamazaki, Time-resolved total internal reflection fluorescence spectroscopy for surface photophysics studies, J. Phys. Chem. 90, 5830-5835 (1986). 

A. Itaya, T. Yamada, K. Tokuda, and H. Masuhara, Interfacial characteristics of poly(methyl methacrylate) film Aggregation of pyrene and micropolarity revealed by time-resolved total internal reflection fluorescence spectroscopy, Polym. J. 22, 697-704 (1990). 

Picosecond time-resolved total internal reflection fluorescence spectroscopy was applied to analyze the proton-transfer reaction of INpOH in water-sapphire interface layers [206], The rate constant of the proton-transfer reaction from excited neutral species became slow in the interface layer as compared with that in the bulk aqueous solution and decreased smoothly with increasing penetration depth in the interfacial layer up to 100 nm. The anomaly was interpreted in terms of rotational fluctuations of water aggregates in the interface layer. 

Among the other spectroscopies, either in the direct or reflection mode, fluorescence spectroscopy may be mentioned. A promising variant is Total Internal Reflection Fluorescence spectroscopy (TIRF). The decay rate of an excited fluorescing probe is usually Interpreted in terms of the local fluidity and polarity. The technique has been used to estimate the extent of ordering inside adsorbed surfactant layers, but this is not an absolute method because a fluorescent probe has to be inserted, and such probes themselves affect the local fluidity. More rigorous are fluorescence experiments with molecules that possess such a probe as an intrinsic part of their structure, such as tryptophans in... 

Total internal reflection fluorescence spectroscopy has been used to assay the fluorescence of tryptophan in proteins or of fluorescence markers. Morphine has been determined in this manner with a detection limit of 0.2 pmol/l on a quartz support bearing immobilized fluorescein-labeled antihapten (Kronick and Little, 1973). 

TIME-RESOLVED TOTAL INTERNAL REFLECTION FLUORESCENCE SPECTROSCOPY... 

Fig. 1. Optical set for total internal reflection fluorescence spectroscopy. 6 incident angle of an excitation laser beam. S-film thin surface film. B-film thick bulk film.

A Comparative Study Using Total Internal Reflection Fluorescence Spectroscopy and Molecular Graphics... 

The application of total internal reflection fluorescence spectroscopy (TIRF) by this laboratory to the study of protein adsorption at solid-liquid interfaces is reviewed. TIRF has been used to determine adsorption isotherms and adsorption rates from single-and multi-component protein solutions. Initial adsorption rates of BSA can be explained qualitatively by the properties of the adsorbing surface. Most recently, a TIRF study using monoclonal antibodies to probe the conformation of adsorbed sperm whale myoglobin (Mb) elucidated two aspects of the Mb adsorption process 1) Mb adsorbs in a non-random manner. 2) Conformational changes of adsorbed Mb, if they occur, are minor and confined to local regions of the molecule. Fluorescence energy transfer and proteolytic enzyme techniques, when coupled with TIRF, can characterize, respectively, the conformation and orientation of adsorbed Mb. 

Absorption Linear Dichroism and Total Internal Reflection Fluorescence Spectroscopies. 346... 

Applying attenuated total reflection Fourier transform infrared and total internal reflection fluorescence spectroscopy, the same group studied the secondary structure and aggregation properties of different proteins that are adsorbed onto planar PAA brushes (Hollmann, Steitz, Czeslik, 2008 Reichhart Czeslik, 2008). They found... 

The surface plasmon related optical enhancements at resonance are in sharp contrast to the maximum value one might find at the critical angle c for a mere total internal reflection geometry displayed in Figure 3(c). Here, the coherent superposition of the incoming and the outgoing light beam results in an enhancement of a factor of 4 only However, this has been utilized in the past for a surface-specific fluorescence technique, i.e., the total internal reflection fluorescence spectroscopy (TIRF) ... 

T. Yamashita, T. Uchida, T. Fukushima, andN. Teramae,/. Phys. Chem. B, 107,4786 (2003). Solvation Dynamics of Fluorophores with an Anthroyloxy Group at the Heptane/Water Interface as Studied by Time-Resolved Total Internal Reflection Fluorescence Spectroscopy. 

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