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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]
The bleached profile will not be a radial step function. If the bleached spot is effectively a single pixel then the bleaching as a function of position will typically be diffraction limited and determined by the optics of the confocal laser scanning microscope used. This is not a radial step function and also varies along the axis ...
Commonly, FCS is employed in the context of optical microscopy, in particular confocal microscopy or two-photon excitation microscopy. In these techniques light is focused on a sample and the measured fluorescence intensity fluctuations (due to diffusion , physical or chemical reactions, aggregation, etc.) are analyzed using the temporal ...
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the photon emission of a fluorophore from a sample. It can be used as an imaging technique in confocal microscopy , two-photon excitation microscopy , and multiphoton tomography.
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 ...
Schematic and SEM images of a conventional SThM tip based on an Au–Cr thermocouple. [1] SThM using the N-V center in diamond. (a) Schematics of experimental setup. An electric current is applied to the arms of an AFM cantilever (phosphorus-doped Si, P:Si) and heats up the end section above the tip (intrinsic Si, i-Si).
Colocalization is used in real-time single-molecule fluorescence microscopy to detect interactions between fluorescently labeled molecular species. In this case, one species (e.g. a DNA molecule) is typically immobilized on the imaging surface, and the other species (e.g. a DNA-binding protein) is supplied to the solution.
Raman microscopy, and in particular confocal microscopy, can reach down to sub-micrometer lateral spatial resolution. [7] Because a Raman microscope is a diffraction-limited system, its spatial resolution depends on the wavelength of light and the numerical aperture of the focusing element. In confocal Raman microscopy, the diameter of the ...
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