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[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]
Multicolor fluorescence image of living HeLa cells. Fluorescence imaging is a type of non-invasive imaging technique that can help visualize biological processes taking place in a living organism. Images can be produced from a variety of methods including: microscopy, imaging probes, and spectroscopy.
A 4Pi microscope is a laser-scanning fluorescence microscope with an improved axial resolution. The typical value of 500–700 nm can be improved to 100–150 nm, which corresponds to an almost spherical focal spot with 5–7 times less volume than that of standard confocal microscopy.
From the beginning of the process, STED has allowed fluorescence microscopy to perform tasks that had been only possible using electron microscopy. As an example, STED was used for the elucidation of protein structure analysis at a sub-organelle level. The common proof of this level of study is the observation of cytoskeletal filaments.
Photo-activated localization microscopy (PALM or FPALM) [1] [2] and stochastic optical reconstruction microscopy (STORM) [3] are widefield (as opposed to point scanning techniques such as laser scanning confocal microscopy) fluorescence microscopy imaging methods that allow obtaining images with a resolution beyond the diffraction limit.
With no modification to the microscope, i.e. with a simple wide field light microscope, the quality of optical sectioning is governed by the same physics as the depth of field effect in photography. For a high numerical aperture lens, equivalent to a wide aperture , the depth of field is small ( shallow focus ) and gives good optical sectioning.
Antonie van Leeuwenhoek (1632–1723). The field of microscopy (optical microscopy) dates back to at least the 17th-century.Earlier microscopes, single lens magnifying glasses with limited magnification, date at least as far back as the wide spread use of lenses in eyeglasses in the 13th century [2] but more advanced compound microscopes first appeared in Europe around 1620 [3] [4] The ...
Bright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light , and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample.
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