Confocal microscopy is an optical imaging technique used to increase micrograph contrast and/or to reconstruct three-dimensional images by using a spatial pinhole to eliminate out-of-focus light or flare in specimens that are thicker than the focal plane. A micrograph, microphotograph or photomicrograph is a Photograph or similar image taken through a Microscope or similar device to show Contrast is the difference in visual properties that makes an object (or its representation in an image distinguishable from other objects and the background In the History of the United States, " Reconstruction " refers to the time between 1863 and 1877 when the U An image (from Latin imago) or picture is an artifact usually two-dimensional that has a similar appearance to some subject &mdashusually Lens flare is the light scattered in lens systems through generally unwanted image formation mechanisms such as internal reflection and Scattering from The cardinal points and the associated cardinal planes are a set of special points and planes in an optical system which help in the analysis [1] This technique has been gaining popularity in the scientific and industrial communities. Typical applications include life sciences and semiconductor inspection. Foundations of modern biology There are five unifying principles A semiconductor' is a Solid material that has Electrical conductivity in between a conductor and an insulator; it can vary over that
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Basic concept
The principle of confocal imaging was patented by Marvin Minsky in 1957. Marvin Lee Minsky (born August 9, 1927) is an American cognitive scientist in the field of Artificial intelligence (AI co-founder [2] In a conventional (i. e. , wide-field) fluorescence microscope, the entire specimen is flooded in light from a light source. A fluorescence microscope (colloquially synonymous with epifluorescent microscope) is a light Microscope used to study properties of organic or inorganic substances In Biology, a specimen is an individual Animal, part of an animal Plant, part of a plant or Microorganism used as a representative to Due to the conservation of light intensity transportation, all parts of the specimen throughout the optical path will be excited and the fluorescence detected by a photodetector or a camera. In Physics, intensity is a measure of the time-averaged Energy Flux. Photosensors or photodetectors are Sensors of Light or other Electromagnetic energy A camera is a device used to capture images either as still Photographs or as sequences of moving images ( Movies or Videos. In contrast, a confocal microscope uses point illumination and a pinhole in an optically conjugate plane in front of the detector to eliminate out-of-focus information. Only the light within the focal plane can be detected, so the image quality is much better than that of wide-field images. As only one point is illuminated at a time in confocal microscopy, 2D or 3D imaging requires scanning over a regular raster (i. e. a rectangular pattern of parallel scanning lines) in the specimen. The thickness of the focal plane is defined mostly by the square of the numerical aperture of the objective lens, and also by the optical properties of the specimen and the ambient index of refraction. In Optics, the numerical aperture ( NA) of an optical system is a Dimensionless number that characterizes the range of angles over which the system can accept An objective in Optics is the lens or Mirror in a Microscope, Telescope, camera or other optical instrument
Types
Three types of confocal microscopes are commercially available: Confocal laser scanning microscopes, spinning-disk (Nipkow disk) confocal microscopes and Programmable Array Microscopes (PAM). Confocal laser scanning microscopy ( CLSM or LSCM) is a technique for obtaining high- resolution optical images A Nipkow disk (sometimes Anglicized as Nipkov disk also known as scanning disk, is a mechanical geometrically operating Image scanning device invented by Confocal laser scanning microscopy yields better image quality than Nipkow and PAM, but the imaging frame rate is very slow (less than 3 frames/second); spinning-disk confocal microscopes can achieve video rate imaging—a desirable feature for dynamic observations such as live cell imaging. Frame rate, or frame frequency, is the measurement of the Frequency (rate at which an imaging device produces unique consecutive images called frames The
References
- ^ Pawley JB (editor) (2006). Handbook of Biological Confocal Microscopy, 3rd ed. , Berlin: Springer. ISBN 038725921x.
- ^ US patent 3013467
External links
- Molecular Expressions: Laser Scanning Confocal Microscopy
- Live Cell Confocal Microscopy. Live Cell Confocal Microscopy from Andor Technolgy
- Leica MicroscopyU.
Comprehensive introduction to confocal microscopy.
- Nikon's MicroscopyU. Comprehensive introduction to confocal microscopy.
- Emory’s Physics Department. Introduction to confocal microscopy and fluorescence.
- The Science Creative Quarterly's overview of confocal microscopy - high res images also available.
- Programmable Array Microscope - Confocal Microscope Capabilities.
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