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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]
Scanning laser ophthalmoscopy developed as a method to view a distinct layer of the living eye at the microscopic level. The use of confocal methods to diminish extra light by focusing detected light through a small pinhole made possible the imaging of individual layers of the retina with greater distinction than ever before. [4]
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.
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 ...
Confocal microscopy uses a scanning point or points of light to illuminate the sample. In conjunction with a pinhole at a conjugate focal plane this acts to filter out light from sources outside the focal plane to improve optical sectioning.
Edge filters are used for removal of all primary laser light. Resolution as low as 10 nm can be achieved using this technique. [citation needed] Near field infrared spectrometry and near-field dielectric microscopy [19] use near-field probes to combine sub-micron microscopy with localized IR spectroscopy. [25]
A 4Pi microscope is a laser scanning fluorescence microscope with an improved axial resolution.With it the typical range of the axial resolution 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.
Photoacoustic imaging or optoacoustic imaging is a biomedical imaging modality based on the photoacoustic effect.Non-ionizing laser pulses are delivered into biological tissues and part of the energy will be absorbed and converted into heat, leading to transient thermoelastic expansion and thus wideband (i.e. MHz) ultrasonic emission.