Total internal reflection fluorescence microscopeA total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide. The technique is based on the principle that when excitation light is totally internally reflected in a transparent solid coverglass at its interface with a liquid medium, an electromagnetic field, also known as an evanescent wave, is generated at the solid-liquid interface with the same frequency as the excitation light.
Near-field scanning optical microscopeNear-field scanning optical microscopy (NSOM) or scanning near-field optical microscopy (SNOM) is a microscopy technique for nanostructure investigation that breaks the far field resolution limit by exploiting the properties of evanescent waves. In SNOM, the excitation laser light is focused through an aperture with a diameter smaller than the excitation wavelength, resulting in an evanescent field (or near-field) on the far side of the aperture.
Optical microscopeThe optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast. The object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope.
Confocal microscopyConfocal 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. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object.
MicroscopyMicroscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection, or refraction of electromagnetic radiation/electron beams interacting with the specimen, and the collection of the scattered radiation or another signal in order to create an image.
Virtual imageIn optics, an image is defined as the collection of focus points of light rays coming from an object. A is the collection of focus points made by converging rays, while a virtual image is the collection of focus points made by extensions of diverging rays. In other words, a virtual image is found by tracing real rays that emerge from an optical device (lens, mirror, or some combination) backward to perceived or apparent origins of ray divergences.
Oil immersionIn light microscopy, oil immersion is a technique used to increase the resolving power of a microscope. This is achieved by immersing both the objective lens and the specimen in a transparent oil of high refractive index, thereby increasing the numerical aperture of the objective lens. Without oil, light waves reflect off the slide specimen through the glass cover slip, through the air, and into the microscope lens (see the colored figure to the right).
Inverted microscopeAn inverted microscope is a microscope with its light source and condenser on the top, above the stage pointing down, while the objectives and turret are below the stage pointing up. It was invented in 1850 by J. Lawrence Smith, a faculty member of Tulane University (then named the Medical College of Louisiana). The stage of an inverted microscope is usually fixed, and focus is adjusted by moving the objective lens along a vertical axis to bring it closer to or further from the specimen.
MagnificationMagnification is the process of enlarging the apparent size, not physical size, of something. This enlargement is quantified by a size ratio called optical magnification. When this number is less than one, it refers to a reduction in size, sometimes called de-magnification. Typically, magnification is related to scaling up visuals or s to be able to see more detail, increasing resolution, using microscope, printing techniques, or digital processing. In all cases, the magnification of the image does not change the perspective of the image.
Super-resolution microscopySuper-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that use the Pendry Superlens and near field scanning optical microscopy) or on the far-field.
Fluorescence microscopeA fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. "Fluorescence microscope" refers to 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.
Magnifying glassA magnifying glass is a convex lens that is used to produce a magnified of an object. The lens is usually mounted in a frame with a handle. A magnifying glass can be used to focus light, such as to concentrate the sun's radiation to create a hot spot at the focus for fire starting. A sheet magnifier consists of many very narrow concentric ring-shaped lenses, such that the combination acts as a single lens but is much thinner. This arrangement is known as a Fresnel lens.
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.
Cell fractionationIn cell biology, cell fractionation is the process used to separate cellular components while preserving individual functions of each component. This is a method that was originally used to demonstrate the cellular location of various biochemical processes. Other uses of subcellular fractionation is to provide an enriched source of a protein for further purification, and facilitate the diagnosis of various disease states. Tissue is typically homogenized in a buffer solution that is isotonic to stop osmotic damage.
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.
Curved mirrorA curved mirror is a mirror with a curved reflecting surface. The surface may be either convex (bulging outward) or concave (recessed inward). Most curved mirrors have surfaces that are shaped like part of a sphere, but other shapes are sometimes used in optical devices. The most common non-spherical type are parabolic reflectors, found in optical devices such as reflecting telescopes that need to image distant objects, since spherical mirror systems, like spherical lenses, suffer from spherical aberration.
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.
SuperlensA superlens, or super lens, is a lens which uses metamaterials to go beyond the diffraction limit. The diffraction limit is a feature of conventional lenses and microscopes that limits the fineness of their resolution depending on the illumination wavelength and the numerical aperture NA of the objective lens. Many lens designs have been proposed that go beyond the diffraction limit in some way, but constraints and obstacles face each of them. In 1873 Ernst Abbe reported that conventional lenses are incapable of capturing some fine details of any given image.
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.
Super-resolution imagingSuper-resolution imaging (SR) is a class of techniques that enhance (increase) the of an imaging system. In optical SR the diffraction limit of systems is transcended, while in geometrical SR the resolution of digital is enhanced. In some radar and sonar imaging applications (e.g. magnetic resonance imaging (MRI), high-resolution computed tomography), subspace decomposition-based methods (e.g. MUSIC) and compressed sensing-based algorithms (e.g., SAMV) are employed to achieve SR over standard periodogram algorithm.
