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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]
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 ...
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 ...
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.
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 ...
Similar to confocal microscopy, the laser in CLE filtered by the pinhole excites the fluorescent dye through a beam splitter and objective lens. The fluorescent emission then follows similar paths into the detector. A pinhole is used to select emissions from the desired focal plane. Two categories of CLE exist, namely probe-based (pCLE) and the ...
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.
Lattice light-sheet microscopy is a modified version of light sheet fluorescence microscopy that increases image acquisition speed while decreasing damage to cells caused by phototoxicity. This is achieved by using a structured light sheet to excite fluorescence in successive planes of a specimen, generating a time series of 3D images which can ...