Sphingolipids are a class of lipids containing a backbone of sphingoid bases, a set ofaliphatic amino alcohols that includes sphingosine.

They were discovered in brain extracts in the 1870s and were named for the mythological Sphinx because of their enigmatic nature.[1]These compounds play important roles in signal transmission and cell recognition.Sphingolipidoses, or disorders of sphingolipid metabolism, have particular impact on neural tissue. A sphingolipid with an R group consisting of a hydrogen atom only is a ceramide. Other common R groups include phosphocholine, yielding a sphingomyelin, and various sugar monomers or dimers, yielding cerebrosides and globosides, respectively. Cerebrosides and globosides are collectively known as glycosphingolipids. Structure The long-chain bases, sometimes simply known as sphingoid bases, are the first non-transient products of de novo sphingolipid synthesis in both yeast and mammals. These compounds, specifically known as phytosphingosine and dihydrosphingosine (also known as sphinganine,[2] although this term is less common), are mainly C18 compounds, with somewhat lower levels of C20 bases.[3]Ceramides and glycosphingolipids are N-acyl derivatives of these compounds.[4] The sphingosine backbone is O-linked to a (usually) charged head group such as ethanolamine, serine, or choline. The backbone is also amide-linked to an acyl group, such as a fatty acid. Types Simple sphingolipids, which include the sphingoid bases and ceramides, make up the early products of the sphingolipid synthetic pathways.

Sphingoid bases are the fundamental building blocks of all sphingolipids. The main mammalian sphingoid bases are dihydrosphingosine and sphingosine, while dihydrosphingosine and phytosphingosine are the principle sphingoid bases in yeast.[5][6]Sphingosine, dihydrosphingosine, and phytosphingosine may be phosphorylated. Ceramides, as a general class, are N-acylated sphingoid bases lacking additional head groups. Dihydroceramide is produced by N-acylation of dihydrosphingosine. Dihydroceramide is found in both yeast and mammalian systems.

Ceramide is produced in mammalian systems by desaturation of dihydroceramide by dihydroceramide desaturase 1 (DES1). This highly bioactive molecule may also be phosphorylated to form ceramide-1phosphate. Phytoceramide is produced in yeast by hydroxylation of dihydroceramide at C-4. Complex sphingolipids may be formed by addition of head groups to ceramide or phytoceramide:

Sphingomyelins have a phosphocholine or phosphoethanolamine molecule with an ester linkage to the 1-hydroxy group of a ceramide. Glycosphingolipids are ceramides with one or more sugar residues joined in a -glycosidic linkage at the 1-hydroxyl position (see image). Cerebrosides have a single glucose or galactose at the 1-hydroxy position. Sulfatides are sulfated cerebrosides. Gangliosides have at least three sugars, one of which must be sialic acid. Inositol-containing ceramides, which are derived from phytoceramide, are produced in yeast. These include inositol phosphorylceramide, mannose inositol phosphorylceramide, and mannose diinositol phosphorylceramide.

Transmission electron microscope (TEM) The original form of electron microscope, the transmission electron microscope (TEM) uses a high voltage electron beam to create an image. The electrons are emitted by an electron gun, commonly fitted with a tungsten filament cathode as the electron source. The electron beam is accelerated by an anode typically at +100 keV (40 to 400 keV) with respect to the cathode, focused by electrostaticand electromagnetic lenses, and transmitted through the specimen that is in part transparent to electrons and in part scatters them out of the beam. When it emerges from the specimen, the electron beam carries information about the structure of the specimen that is magnified by the objective lens system of the microscope. The spatial variation in this information (the "image") may be viewed by projecting the magnified electron image onto a fluorescent viewing screen coated with a phosphor or scintillator material such as zinc sulfide. Alternatively, the image can be photographically recorded by exposing a photographic film or plate directly to the electron beam, or a high-resolution phosphor may be coupled by means of a lens optical system or a fibre optic light-guide to the sensor of a CCD (charge-

coupled device) camera. The image detected by the CCD may be displayed on a monitor or computer. Resolution of the TEM is limited primarily by spherical aberration, but a new generation of aberration correctors have been able to partially overcome spherical aberration to increase resolution. Hardware correction of spherical aberration for the high-resolution transmission electron microscopy (HRTEM) has allowed the production of images with resolution below 0.5 angstrom (50 picometres)[1]and magnifications above 50 million times.[8] The ability to determine the positions of atoms within materials has made the HRTEM an important tool for nanotechnologies research and development.[9] An important mode of TEM utilization is electron diffraction. The advantages of electron diffraction over X-ray crystallography are that the specimen need not be a single crystal or even a polycrystalline powder, and also that the Fourier transform reconstruction of the object's magnified structure occurs physically and thus avoids the need for solving the phase problem faced by the X-ray crystallographers after obtaining their X-ray diffraction patterns of a single crystal or polycrystalline powder. The major disadvantage of the transmission electron microscope is the need for extremely thin sections of the specimens, typically about 100 nanometers. Biological specimens typically require to be chemically fixed, dehydrated and embedded in a polymer resin to stabilize them sufficiently to allow ultrathin sectioning. Sections of biological specimens, organic polymers and similar materials may require special `staining' with heavy atom labels in order to achieve the required image contrast. Scanning electron microscope Unlike the TEM, where electrons of the high voltage beam carry the image of the specimen, the electron beam of the scanning electron microscope (SEM)[10] does not at any time carry a complete image of the specimen. The SEM produces images by probing the specimen with a focused electron beam that is scanned across a rectangular area of the specimen (raster scanning). When the electron beam interacts with the specimen, it loses energy by a variety of mechanisms. The lost energy is converted into alternative forms such as heat, emission of low-energy secondary electrons and high-energy backscattered electrons, light emission (cathodoluminescence) or X-ray emission, which provide signals carrying information about the properties of the specimen surface, such as its topography and composition. The image displayed by an SEM maps the varying intensity of

any of these signals into the image in a position corresponding to the position of the beam on the specimen when the signal was generated. In the SEM image of an ant shown at right, the image was constructed from signals produced by a secondary electron detector, the normal or conventional imaging mode in most SEMs. Generally, the image resolution of an SEM is about an order of magnitude poorer than that of a TEM. However, because the SEM image relies on surface processes rather than transmission, it is able to image bulk samples up to many centimetres in size and (depending on instrument design and settings) has a great depth of field, and so can produce images that are good representations of the three-dimensional shape of the sample. Another advantage of SEM is its variety called environmental scanning electron microscope (ESEM) can produce images of sufficient quality and resolution with the samples being wet or contained in low vacuum or gas. This greatly facilitates imaging biological samples that are unstable in the high vacuum of conventional electron microscopes. Difference Between Scanning Electron Microscope and Transmission Electron Microscope

An electron microscope produces its ultra fine image by passing a particle beam of electrons through electrostatic or electromagnetic lenses, similar to the principle of a light microscope. However, since the wavelength of an electron beam is so much shorter. A shorter wavelength means a higher resolution. Electron microscopes are a general category in which there are several varieties. The two most common are transmission electron microscopes and scanning electron microscopes. Both use a beam of electrons to view the very small, but the beam acts in different ways.
A transmission electron microscope uses a high-powered beam to essentially shoot electrons through the object. The electron beam first passes through a condenser lens in order to concentrate the beam on the object. Then the beam goes through the object. Some of the electrons pass all the way through; others hit molecules in

the object and scatter. The modified beam then passes through an objective lens, a projector lens and onto a fluorescent screen where the final image is observed. Because the electron beam passes entirely through the object, the pattern of scatter gives the observed a comprehensive view of the interior of the object. A scanning electron microscope doesnt use a concentrated electron beam to penetrate the object, as a transmission electron microscope does. Instead it scans a beam across the object. During the scanning the beam loses energy in different amounts according to the surface it is on. A scanning electron microscope measures the lost energy to create a three-dimensional picture of the surface of an object. While not quite as powerful as a transmission electron microscope, a scanning electron microscope is able to produce comprehensive magnified images of much larger objects, like that of an ant. Recently, other electron microscopes have been developed that combine transmission and scanning technologies. However, all electron microscopes, transmission, scanning or otherwise employ the basic principle of magnifying an object through the use of an electron beam.

Phase contrast microscopy is an optical microscopy illumination technique that convertsphase shifts in light passing through a transparent specimen to brightness changes in the image. The phase shifts themselves are invisible to the human eye, but become visible when they are shown as brightness changes. Phase contrast microscopy is particularly important in biology. Many biological structures can be revealed by it that are not visible with a simpler bright field microscope. These structures were often made visible to earlier microscopists by staining the slide. This requires additional preparation and it also kills the cell. A phase contrast microscope without staining, and with live cells, allowed the in vivo study of the cell cycle. When light travels through a medium other than vacuum, interaction with this medium causes itsamplitude and phase to change in a way which depends on properties of the medium. Changes in amplitude arise from absorption of light,

which is often wavelength dependent and may give rise to colours. The human eye measures only the energy of light arriving on the retina, so changes in phase are not easily observed under optimal bright field illumination, yet often these changes in phase carry much important information. The same situation applies in a typical microscope with "Khler" bright field illumination, i.e., although the phase variations introduced by the sample are preserved by the instrument (at least within the instrumental limits of imaging perfection) this information is lost in the process of image recording, which measures only light intensity. In order to make phase variations observable, it is necessary to combine the light passing through the sample with a reference so that the resulting interference reveals the phase structure of the sample. This problem was first appreciated by Frits Zernike during his study of diffraction gratings. During the course of his work he realised that it is necessary both to achieve interference with a reference beam, and (for maximizing the contrast achieved with the technique) to introduce a phase shift to this reference beam so that the no-phase-change condition gives rise to completely destructive interference. He later realized that the same technique can be applied to optical microscopy. The necessary phase shift is introduced by rings etched accurately onto glass plates so that they introduce the required phase shift when inserted into the optical path of the microscope. When in use, this technique allows the phase of the light passing through the object under study to be inferred from the intensity of the image produced by the microscope. This methodology is known as the phase-contrast technique. In optical microscopy many biological objects such as cell components in protozoans, bacteria and sperm tails are fully transparent unless stained. (Staining is a difficult and time-consuming procedure which can destroy or alter the specimen structure). The difference in densities and composition within the imaged objects however often give rise to changes in the phase of light passing through them, hence they are sometimes called "phase objects". Using the phasecontrast technique makes these structures visible and allows their study in living specimens. This phase contrast technique proved to be such an advancement in microscopy that Zernike was awarded the Nobel prize (physics) in 1953.