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By virtue of the linearity property of optical non-coherent imaging systems, i.e., . Image(Object 1 + Object 2) = Image(Object 1) + Image(Object 2). the image of an object in a microscope or telescope as a non-coherent imaging system can be computed by expressing the object-plane field as a weighted sum of 2D impulse functions, and then expressing the image plane field as a weighted sum of the ...
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 image of a point source is also a three dimensional (3D) intensity distribution which can be represented by a 3D point-spread function. 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).
Optical response from a point source. From left to right: calculated intensity distributions xy (top) and rz (bottom), with logarithmic scale, for a point source imaged by means of a wide field (a), 2PEF (b) and confocal microscopy (c). The 2PEF and confocal forms have a better signal-to-noise ratio than the wide field. The 2PEF distribution is ...
In microscopy, NA is important because it indicates the resolving power of a lens. The size of the finest detail that can be resolved (the resolution) is proportional to λ / 2NA , where λ is the wavelength of the light. A lens with a larger numerical aperture will be able to visualize finer details than a lens with a smaller numerical ...
Because STED selectively deactivates the fluorescence, it can achieve resolution better than traditional confocal microscopy. Normal fluorescence occurs by exciting an electron from the ground state into an excited electronic state of a different fundamental energy level (S0 goes to S1) which, after relaxing back to the vibrational ground state ...
[1] [2] A fluorescence microscope is 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]
In fluorescence microscopy the excitation and emission are typically on different wavelengths. In total internal reflection fluorescence microscopy a thin portion of the sample located immediately on the cover glass is excited with an evanescent field, and recorded with a conventional diffraction-limited objective, improving the axial resolution.