Laser scanning fluorescence microscopy

Strickler J FI and Webb W W 1990 Two-photon excitation in laser scanning fluorescence microscopy Proc. SPIE 13948107-18... 

W. W. (1990) Two-photon laser scanning fluorescence microscopy. Science, 248, 73-76. 

McConnell, G. (2004). Confocal laser scanning fluorescence microscopy with a visible continuum source. Opt. Express 12, 2844—50. 

Multi-spectral imaging and linear unmixing add a whole new dimension to laser scanning fluorescence microscopy. Biotechniques 31, 1272-8. 

Two-photon excitation provides intrinsic 3-D resolution in laser scanning fluorescence microscopy. The 3-D sectioning effect is comparable to that of confocal microscopy, but it offers two advantages with respect to the latter because the illumination is concentrated in both time and space, there is no out-of-focus photo-bleaching, and the excitation beam is not attenuated by out-of-focus absorption, which results in increased penetration depth of the excitation light. 

Confocal laser scanning fluorescence microscopy was used to study the exposure of the avidin-specific binding sites in the Av-GEB platform by the immobilization of a small and flexible biotinylated fluorescein molecule as a fluorescence marker. Fluorescence microscopy thus confirms that Av-GEB platform exposes active binding sites for biotin, acting as affinity matrix (Fig. 21.2B). After use, the electrode surface can be renewed by a simple polishing procedure for further uses, highlighting a clear advantage of this new material with respect to surface-modified approaches such as classical biosensors and other common... 

Other methods of imaging fat crystals and fat crystal networks (not all of them optical) include confocal laser scanning fluorescence microscopy, multiple photon microscopy, atomic force microscopy and electron microscopy (Narine and Marangoni, 1999). 

In recent years, many other techniques have been employed to elucidate the structure of fat crystal networks including confocal laser scanning fluorescence microscopy (Heertje et al. 1987) and multiple photon microscopy (Marangoni and Hartel 1996). Another advance has been the development of three-dimensional imaging. 

Denk W, Strickler JH. Webb WW (1990) 2-Photon laser scanning fluorescence microscopy. Science 248 73-... 

Confocal laser-scanning fluorescence microscopy (CLSM). The micrographs were obtained by means of a Leica confocal scanning system mounted to a Leica Aristoplan. A 1 OOX oil immersion objective with the numerical aperture 1.4 was used. The standard filter settings for fluorescent excitation and emission were used. 

Guthoff RF, Wienss H, Hahnel C et al (2005) Epithelial innervation of human cornea. A three-dimensional study using confocal laser scanning fluorescence microscopy. Cornea 24 608-13... 

The latter process was performed by immersing the patterned amino polymer films into a solution containing FITC, then the film was intensively washed, and the resulting fluorescent structure finally was visualized with a confocal laser scanning fluorescence microscopy. The result is displayed in figure 3b). 

With the advent of confocal laser scanning fluorescence microscopy (CLSFM) [39], multiple photon microscopy (MPM) [40,41], and atomic force microscopy (AFM) [5], three new tools have been added to the standard tools of light microscopy (LM) [42-45] and electron microscopy (EM) [35,46-50] that were most widely used in the past to study the microstructure of fats and foods in general. The work of Heertje et al. [35,36,39,51] on visualization of the microstructure in fats remains one of the most important contributions to the field. In their method, a cold solvent mixture (butanol-methanol) was used to remove the liquid oil form the solid fat in a sample mounted on a special holder. After removal of the liquid oil, the structure of the solid fat network could be visualized. 

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