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The observation of sub-wavelength structures with microscopes is difficult because of the Abbe diffraction limit.Ernst Abbe found in 1873, [2] and expressed as a formula in 1882, [3] that light with wavelength , traveling in a medium with refractive index and converging to a spot with half-angle will have a minimum resolvable distance of
There is a diffraction-limited resolution depending on incident wavelength; in visible range, the resolution of optical microscopy is limited to approximately 0.2 micrometres (see: microscope) and the practical magnification limit to ~1500x.
All optical microscopes are diffraction-limited because of the wave nature of light. Current research focuses on techniques to go beyond this limit known as the Rayleigh criterion. The use of SIL can achieve spatial resolution better than the diffraction limit in air, for both far-field imaging [3] [4] and near-field imaging.
However, resolution below this theoretical limit can be achieved using super-resolution microscopy. These include optical near-fields (Near-field scanning optical microscope) or a diffraction technique called 4Pi STED microscopy. Objects as small as 30 nm have been resolved with both techniques.
Confocal microscopes work on the principle of point excitation in the specimen (diffraction limited spot) and point detection of the resulting fluorescent signal. A pinhole at the detector provides a physical barrier that blocks out-of-focus fluorescence. Only the in-focus, or central spot of the Airy disk, is recorded.
Diffraction can also be a concern in some technical applications; it sets a fundamental limit to the resolution of a camera, telescope, or microscope. Other examples of diffraction are considered below.
STED microscopy is one of several types of super resolution microscopy techniques that have recently been developed to bypass the diffraction limit of light microscopy to increase resolution. STED is a deterministic functional technique that exploits the non-linear response of fluorophores commonly used to label biological samples in order to ...
The ability of a lens to resolve detail is usually determined by the quality of the lens, but is ultimately limited by diffraction.Light coming from a point source in the object diffracts through the lens aperture such that it forms a diffraction pattern in the image, which has a central spot and surrounding bright rings, separated by dark nulls; this pattern is known as an Airy pattern, and ...