Two-photon excitation microscopyTwo-photon excitation microscopy (TPEF or 2PEF) is a fluorescence imaging technique that is particularly well-suited to image scattering living tissue of up to about one millimeter in thickness. Unlike traditional fluorescence microscopy, where the excitation wavelength is shorter than the emission wavelength, two-photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. The laser is focused onto a specific location in the tissue and scanned across the sample to sequentially produce the image.
Papanicolaou stainPapanicolaou stain (also Papanicolaou's stain and Pap stain) is a multichromatic (multicolored) cytological staining technique developed by George Papanicolaou in 1942. The Papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. Although most notable for its use in the detection of cervical cancer in the Pap test or Pap smear, it is also used to stain non-gynecological specimen preparations from a variety of bodily secretions and from small needle biopsies of organs and tissues.
DiffractionDiffraction is the interference or bending of waves around the corners of an obstacle or through an aperture into the region of geometrical shadow of the obstacle/aperture. The diffracting object or aperture effectively becomes a secondary source of the propagating wave. Italian scientist Francesco Maria Grimaldi coined the word diffraction and was the first to record accurate observations of the phenomenon in 1660.
Image editingImage editing encompasses the processes of altering s, whether they are digital photographs, traditional photo-chemical photographs, or illustrations. Traditional analog image editing is known as photo retouching, using tools such as an airbrush to modify photographs or editing illustrations with any traditional art medium. Graphic software programs, which can be broadly grouped into vector graphics editors, raster graphics editors, and 3D modelers, are the primary tools with which a user may manipulate, enhance, and transform images.
LightLight or visible light is electromagnetic radiation that can be perceived by the human eye. Visible light is usually defined as having wavelengths in the range of 400–700 nanometres (nm), corresponding to frequencies of 750–420 terahertz, between the infrared (with longer wavelengths) and the ultraviolet (with shorter wavelengths). In physics, the term "light" may refer more broadly to electromagnetic radiation of any wavelength, whether visible or not. In this sense, gamma rays, X-rays, microwaves and radio waves are also light.
Fresnel diffractionIn optics, the Fresnel diffraction equation for near-field diffraction is an approximation of the Kirchhoff–Fresnel diffraction that can be applied to the propagation of waves in the near field. It is used to calculate the diffraction pattern created by waves passing through an aperture or around an object, when viewed from relatively close to the object. In contrast the diffraction pattern in the far field region is given by the Fraunhofer diffraction equation. The near field can be specified by the Fresnel number, F, of the optical arrangement.
Fraunhofer diffractionIn optics, the Fraunhofer diffraction equation is used to model the diffraction of waves when plane waves are incident on a diffracting object, and the diffraction pattern is viewed at a sufficiently long distance (a distance satisfying Fraunhofer condition) from the object (in the far-field region), and also when it is viewed at the focal plane of an imaging lens. In contrast, the diffraction pattern created near the diffracting object and (in the near field region) is given by the Fresnel diffraction equation.
Diffraction gratingIn optics, a diffraction grating is an optical grating with a periodic structure that diffracts light into several beams travelling in different directions (i.e., different diffraction angles). The emerging coloration is a form of structural coloration. The directions or diffraction angles of these beams depend on the wave (light) incident angle to the diffraction grating, the spacing or distance between adjacent diffracting elements (e.g., parallel slits for a transmission grating) on the grating, and the wavelength of the incident light.
Dark-field microscopyDark-field microscopy (also called dark-ground microscopy) describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. As a result, the field around the specimen (i.e., where there is no specimen to scatter the beam) is generally dark. In optical microscopes a darkfield condenser lens must be used, which directs a cone of light away from the objective lens. To maximize the scattered light-gathering power of the objective lens, oil immersion is used and the numerical aperture (NA) of the objective lens must be less than 1.
LensA lens is a transmissive optical device that focuses or disperses a light beam by means of refraction. A simple lens consists of a single piece of transparent material, while a compound lens consists of several simple lenses (elements), usually arranged along a common axis. Lenses are made from materials such as glass or plastic and are ground, polished, or molded to the required shape. A lens can focus light to form an , unlike a prism, which refracts light without focusing.
Thin lensIn optics, a thin lens is a lens with a thickness (distance along the optical axis between the two surfaces of the lens) that is negligible compared to the radii of curvature of the lens surfaces. Lenses whose thickness is not negligible are sometimes called thick lenses. The thin lens approximation ignores optical effects due to the thickness of lenses and simplifies ray tracing calculations. It is often combined with the paraxial approximation in techniques such as ray transfer matrix analysis.
MicroscopeA microscope () is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a microscope. There are many types of microscopes, and they may be grouped in different ways.
