TIRFM total internal reflection fluorescence

TIRFM total internal reflection fluorescence microscopy TLM thermal lens microscopy... 

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...

To discriminate between surface bound protein molecules and those in bulk solution, total internal reflection fluorescence microscopy (TIRFM)41 55 was employed. TIRFM creates an evanescence wave that decays as a function of distance from the surface as ... 

Figure 2.6 Application of splinted RNA ligation procedure single molecule FRET. (A) Diagram of prism-type total internal reflection fluorescence microscope (TIRFM) for single molecule FRET measurements. (B) Distribution of single-molecule FRET values for dye-labeled telomerase RNA molecules generated by splinted RNA ligation. (C) Dye intensity and FRET traces of a single telomerase RNA molecule Cy3 emission (green), Cy5 emission (red), FRET ratio (blue).

Complementary microscopy techniques can be used to follow the morphology and growth of fibrils either on a surface or in aliquots taken from the assembly solution including total internal reflection fluorescence microscopy (TIRFM) (Ban et al., 2004), transmission electron microscopy (TEM), or atomic force microscopy (AFM). 

Fig. 15.1. Schematic representation of amyloid fibrils revealed by total internal reflection fluorescence microscopy, (a) The penetration depth of the evanescent field formed by the total internal reflection of laser light is 150nm for a laser light at 455 nm, so only amyloid fibrils lying parallel to the slide glass surface were observed. (b) Schematic diagram of a prism-type TIRFM system on an inverted microscope. ISIT image-intensifier-coupled silicone intensified target camera, CCD charge-coupled device camera...

The use of an evanescent wave to excite fluorophores selectively near a solid-fluid interface is the basis of the technique total internal reflection fluorescence (TIRF). It can be used to study theadsorption kinetics of fluorophores onto a solid surface, and for the determination of orientational order and dynamics in adsorption layers and Langmuir-Blodgett films. TIRF microscopy (TIRFM) may be combined with FRAP ind FCS measurements to yield information about surface diffusion rates and the formation of surface aggregates. 

Total Internal Reflection Fluorescence Microscopy (TIRFM) is another useful tool for studying the reactions of individual molecules adsorbed, adhered or bound to surfaces. Typical applications are membrane fusion of vesicles [177], conformational and orientation changes [178] and lateral mobility of molecules [179],... 

Total internal reflection fluorescence microscopy (TIRFM) is a promising alternative approach to low background fluorescence imaging [68], For excitation of molecules on a surface or within a thin slice of the sample, an evanescent optical field is used traveling along the interface between a medium with a high refractive index n, (typically a quartz glass prism) and... 

Total Internal Reflection Fluorescence Microscopy (TIRFM)... 

Nishida, S., Funabashi, Y, and Ikai, A. 2002. Combination of AFM with an objective-type total internal reflection fluorescence microscope (TIRFM) for nanomanipulation of single cells. 

TIRFM TIRM Total internal reflection fluorescence (microscopy)... 

Kihm KD, Banerjee A, Choi CK, Takagi T (2004) Near-wall hindered Brownian diffusion of nanoparticles examined by three-dimensional ratiometric total internal reflection fluorescence microscopy (3-D R-TIRFM). Exp Fluid 37 811-824... 

Total internal reflection fluorescence microscopy (TIRFM), or simply TIRF, is the application of fluorescence imaging to evanescent-wave microscopy. The material of interest is tagged with fluorescent material that emits light at wavelengths greater than the excitation photons from the evanescent field, which additionally improves the detectability of the evanescent-wave microscopy technique. 

Figure 3.23 shows the layout of a simplified total internal reflection fluorescence microscope (TIRFM). We briefly tour the configmation here, before expanding on experimental details such as alignment, laser power, and optomechanical considerations. The instrument we describe is suitable for single molecule... 

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