Total internal reflection fluorescence

Protein adsorption has been studied with a variety of techniques such as ellipsome-try [107,108], ESCA [109], surface forces measurements [102], total internal reflection fluorescence (TIRE) [103,110], electron microscopy [111], and electrokinetic measurement of latex particles [112,113] and capillaries [114], The TIRE technique has recently been adapted to observe surface diffusion [106] and orientation [IIS] in adsorbed layers. These experiments point toward the significant influence of the protein-surface interaction on the adsorption characteristics [105,108,110]. A very important interaction is due to the hydrophobic interaction between parts of the protein and polymeric surfaces [18], although often electrostatic interactions are also influential [ 116]. Protein desorption can be affected by altering the pH [117] or by the introduction of a complexing agent [118]. 

Tokunaga M, Kitamura K, Saito K, Iwane A H and Yanagida T 1997 Single molecule imaging of fluorophores and enzymatic reactions achieved by objective-type total internal reflection fluorescence microscopy Biochem. Biophys. Res. Commun. 235 47-53... 

Rotational dynamics of a fluorescent dye adsorbed at the interface provides useful information concerning the rigidity of the microenvironment of liquid-liquid interfaee in terms of the interfacial viscosity. The rotational relaxation time of the rhodamine B dye was studied by time-resolved total internal reflection fluorescent anisotropy. In-plane... 

Jones MA, Bohn PW (2001) Total internal reflection fluorescence and electrocapillary investigations of adsorption at the water-dichloroethane electrochemical interface. 2. Fluorescence-detected linear dichroism investigation of adsorption-driven reorientation of di-N-butylaminonaphthylethenylpyridiniumpropylsulfonate. J Phys Chem B 105(11) 2197-2204... 

Recently, a formalism has been developed to determine the second and the fourth order parameters of films using polarized total internal reflection fluorescence (TIRF) [71]. Similarly to IR-ATR spectroscopy (Section 4), the experiment makes use of p- and s-polarized excitation, but the fluorescence emission (analyzed either in p- or s-direction) is detected normal to the substrate. Two approaches are developed based on the measurements of two intensity ratios. In the first one, the S angle has to be known experimentally or theoretically, and the order parameters (P2) and (P4) can be determined. In the second one, the order parameter (R ) is obtained by another technique, for instance IR-ATR spectroscopy, which allows deducing the order parameter (P4) and (cos2<5). 

Total internal reflection fluorescence lifetime imaging microscopy... 

Axelrod, D. (1981). Cell-substrate contacts illuminated by total internal reflection fluorescence. J. Cell Biol. 89, 141-5. 

Axelrod, D. (2001). Total internal reflection fluorescence microscopy in cell biology. Traffic 2, 764-74. 

Beaumont, V. (2003). Visualizing membrane trafficking using total internal reflection fluorescence microscopy. Biochem. Soc. Trans. 31, 819-23. 

Khakh, B. S., Fisher, J. A., Nashmi, R., Bowser, D. N. and Lester, H. A. (2005). An angstrom scale interaction between plasma membrane ATP-gated P2X2 and alpha4beta2 nicotinic channels measured with fluorescence resonance energy transfer and total internal reflection fluorescence microscopy. J. Neurosci. 25, 6911-20. 

In addition, typical methods of sensing are total internal reflection fluorescence or monitoring of fluorescence resonance energy transfer6,7. The second class is a direct optical detection principle which relies either on reflectometry or refractometry. The latter is connected to evanescent field... 

Fig. 1 Real-time tracking of cell adhesion [42]. (a) Components of a total internal reflection fluorescent microscope (TIRFM). (b) The cell adhesion process (7) a cell approaches the surface, (2) the cell lands, (3) the cell attaches, and (4) the cell spreads out on the surface. The evanescent field was generated by total internal reflection of a laser beam at the glass-water interface. Cells with fluorescently labeled membranes (dashed lines) were plated on SAMs. Cell membranes within the evanescent field (solid line) were observed by TIRFM. Corresponding TIRFM images are shown below...

Axelrod D, Hellen EH, Fulbright RM (1992) Total internal reflection fluorescence. In Lakowicz JR (ed) Topics in fluorescence microscopy. Plenum, New York... 

Burmeister JS, Olivier LA, Reichert WM, Truskey GA (1998) Application of total internal reflection fluorescence microscopy to study cell adhesion to biomaterials. Biomaterials 19 307-325... 

TIRF Total internal reflection fluorescence immunoassay WWTPs Wastewater treatment plants... 

Total internal reflection fluorescence (TIRF) microscopy, fluorescence in situ hybridization (FISH), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM). 

Shino A, Lujioka K, Manabe N, Yamaya S, Goto Y, Yasuhara M, Yamamoto K (2005). Simultaneous multicolor detection system of the single-molecular microbial antigen with total internal reflection fluorescence microscopy. Microbiol. Immunol. 49 461 —170. 

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