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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]
Köhler illumination is a method of specimen illumination used for transmitted and reflected light (trans- and epi-illuminated) optical microscopy.Köhler illumination acts to generate an even illumination of the sample and ensures that an image of the illumination source (for example a halogen lamp filament) is not visible in the resulting image.
An example of an experimentally derived point spread function from a confocal microscope using a 63x 1.4NA oil objective. It was generated using Huygens Professional deconvolution software. Shown are views in xz, xy, yz and a 3D representation. In microscopy, experimental determination of PSF requires sub-resolution (point-like) radiating sources.
As an example, the figure on the right shows the 3D point-spread function in object space of a wide-field microscope (a) alongside that of a confocal microscope (c). Although the same microscope objective with a numerical aperture of 1.49 is used, it is clear that the confocal point spread function is more compact both in the lateral dimensions ...
Diagram illustrating near-field optics, with the diffraction of light coming from NSOM fiber probe, showing wavelength of light and the near-field. [1] Comparison of photoluminescence maps recorded from a molybdenum disulfide flake using NSOM with a campanile probe (top) and conventional confocal microscopy (bottom). Scale bars: 1 μm. [2]
Diagram of a simple microscope. There are two basic types of optical microscopes: simple microscopes and compound microscopes. A simple microscope uses the optical power of a single lens or group of lenses for magnification. A compound microscope uses a system of lenses (one set enlarging the image produced by another) to achieve a much higher ...
[1] [2] "Fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image. [3]
The pupil function or aperture function describes how a light wave is affected upon transmission through an optical imaging system such as a camera, microscope, or the human eye. More specifically, it is a complex function of the position in the pupil [ 1 ] or aperture (often an iris ) that indicates the relative change in amplitude and phase ...