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The success of the phase-contrast microscope has led to a number of subsequent phase-imaging methods. In 1952, Georges Nomarski patented what is today known as differential interference contrast (DIC) microscopy. [8] It enhances contrast by creating artificial shadows, as if the object is illuminated from the side.
Micrasterias furcata imaged in transmitted DIC microscopy Laser-induced optical damage in LiNbO 3 under 150× Nomarski microscopy. Differential interference contrast (DIC) microscopy, also known as Nomarski interference contrast (NIC) or Nomarski microscopy, is an optical microscopy technique used to enhance the contrast in unstained, transparent samples.
With the sample system built, all that is needed is an epifluorescence microscope and a CCD camera to make quantitative intensity measurements. This is a diagram of an example FLIC experimental setup with silicon, three oxide layers and a fluorescently labeled lipid bilayer (the yellow stars represent fluorophores.
Such objects do, however, induce a phase shift that can be observed using a phase contrast microscope. Conventional phase contrast microscopy and related methods, such as differential interference contrast microscopy, visualize phase shifts by transforming phase shift gradients into intensity variations. These intensity variations are mixed ...
The advantages of these methods compared to normal absorption-contrast X-ray imaging is higher contrast for low-absorbing materials (because phase shift is a different mechanism than absorption) and a contrast-to-noise relationship that increases with spatial frequency (because many phase-contrast techniques detect the first or second ...
The contrast between two adjacent areas in a TEM image can be defined as the difference in the electron densities in image plane. Due to the scattering of the incident beam by the sample, the amplitude and phase of the electron wave change, which results in amplitude contrast and phase contrast, correspondingly. Most images have both contrast ...
After its introduction in the 1940s, live-cell imaging rapidly became popular using phase-contrast microscopy. [11] The phase-contrast microscope was popularized through a series of time-lapse movies (see video), recorded using a photographic film camera. [12] Its inventor, Frits Zernike, was awarded the Nobel Prize in 1953. [13]
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]