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Fluorescence and confocal microscopes operating principle. Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. [1]
It uses the technique of confocal laser scanning microscopy for diagnostic imaging of the retina or cornea of the human eye. As a method used to image the retina with a high degree of spatial sensitivity, it is helpful in the diagnosis of glaucoma , macular degeneration , and other retinal disorders.
Laser scanning is the controlled deflection of laser beams, visible or invisible. [1] Scanned laser beams are used in some 3-D printers, in rapid prototyping, in machines for material processing, in laser engraving machines, in ophthalmological laser systems for the treatment of presbyopia, in confocal microscopy, in laser printers, in laser shows, in Laser TV, and in barcode scanners.
Confocal laser scanning microscopy and Two-photon excitation microscopy make use of lasers to obtain blur-free images of thick specimens at various depths. Laser capture microdissection use lasers to procure specific cell populations from a tissue section under microscopic visualization.
Confocal microscope, a widely used variant of epifluorescent illumination that uses a scanning laser to illuminate a sample for fluorescence. Two-photon microscope , used to image fluorescence deeper in scattering media and reduce photobleaching, especially in living samples.
Confocal laser scanning microscopy uses a focused laser beam (e.g. 488 nm) that is scanned across the sample to excite fluorescence in a point-by-point fashion. The emitted light is directed through a pinhole to prevent out-of-focus light from reaching the detector, typically a photomultiplier tube. The image is constructed in a computer ...