Resolution confocal laser scanning

A. Schatzlein and G. Cevc. Non-uniform cellular packing of the stratum corneum and permeability barrier function of intact skin a high-resolution confocal laser scanning microscopy study using highly deformable vesicles (Transfersomes). Br. J. Dermatol. 138 583-592 (1998). 

The classical polarizing light microscope as developed 150 years ago is still the most versatile, least expensive analytical instrument in the hands of an experienced microscopist. Its limitations in terms of resolving power, depth of field, and contrast have been reduced in the last decade, in which we have witnessed a revolution in its evolution. Video microscopy has increased contrast electronically, and thereby revealed structures never before seen. With computer enhancement, unheard of resolutions are possible. There are daily developments in the X-ray, holographic, acoustic, confocal laser scanning, and scanning tunneling micro-... 

Takami S., Getchell M. and Getchell T. (1995). Resolution of sensory and mucoid glycoconjugates with terminal O-galactose residues in the mucomicrovillar complex of the vomeronasal sensory epithelium by dual confocal laser-scanning microscopy. Cell Tiss Res 280, 211-216. 

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 microscopy (CLSM) in conjunction with specific staining techniques is best suited to elucidate intracellular trafficking and localization. CLSM is a specific epifluorescence microscopical technique capable of optical cross-sectioning with a spatial resolution of 1 /urn and below [41, 42],... 

Relatively rapid. Sharp, black, high-resolution reaction product. Sensitive. Permanent preparations. Nontoxic reagents. Can be studied using various forms of microscopy including dark ground, epipolarization, confocal laser scanning and electron microscopy. 

Scanning force microscopy (SFM) has been widely used for visualization of biomedical objects because of combination of extreme resolution, simplicity of sample preparation and ability to operate under physiological conditions. Nowadays SFM is increasingly applied to investigate the ultrastructure of biomedical samples embedded in epoxy resin [1]. In the present work, we are focusing on application of SFM, confocal laser scanning microscopy and ultramicrotomy to the K562 leukemic cells study. 

The combination of SFM, confocal laser scanning microscopy and Ultramicrotomy opens up unique opportunities to visualize external and internal K562 leukemic cellular ultrastructure with a high resolution. 

In summary, we have demonstrated by the presented results that SFM and confocal laser scanning microscopy open unique opportunities to visualize leukemic cells at high resolution. The combined techniques can be used to obtain new more detailed characteristics of leukemic cells than with either technique alone. 

Principles and Characteristics Diffraction limits the spatial resolution of conventional optical microscopy instruments. In practice, the resolution limit is approximately 0.6A., i.e. about 0.5 pm for optical microscopes. Resolution in far-field optical microscopy techniques may be improved (though slightly) by the application of UV (cfr. Chp. 5.3.2) or confocal laser scanning (c/r Chp. 5.3.4). For confocal laser imaging with green light (A. = 500 nm), resolution is limited... 

An alternative to SEM is the use of fluorescence microscopy. The most common technique is confocal laser scanning microscopy (CLSM). CLSM offers two significant advantages over SEM, namely the ability to produce 3D images in situ and the discrimination of species based on fluorescence. Both the x,y resolution and the z penetration depth are to some degree system- and sample-dependant (further discussion of this point can be found in Chapter 4). With the exception of... 

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