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The spectroscopic line shape of the CARS intensity therefore resembles a Fano profile which is shifted with respect to the Raman signal. To compare the spectra from multi-component compounds, the (resonant) CARS spectral amplitude should be compared to the Raman spectral intensity. Coherent anti-Stokes Raman spectrum of microscopy oil.
SRS and CARS were theoretically predicted and experimentally realized in the 1960s. [1] [2] [3] In 1982 the first CARS microscope was demonstrated. [4] In 1999, CARS microscopy using a collinear geometry and high numerical aperture objective were developed in Xiaoliang Sunney Xie's lab at Harvard University. [5]
[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.
PEEM – Photoemission electron microscopy (or photoelectron emission microscopy) PES – Photoelectron spectroscopy; PINEM – photon-induced near-field electron microscopy; PIGE – Particle (or proton) induced gamma-ray spectroscopy, see nuclear reaction analysis; PIXE – Particle (or proton) induced X-ray spectroscopy; PL – Photoluminescence
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]
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.
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 ...
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 ...