42 Savella, Alecza Mae C.

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Darkfield Microscope: A dark field microscope is ideal for viewing objects that are unstained, transparent and absorb little or no light. It is more useful in examining external details, such as outlines, edges, grain boundaries and surface defects than internal structure. Phase-Contrast Microscope: Phase contrast microscopy imparts contrast to unstained biological material by transforming phase differences of light caused by differences in refractive index between cellular components into differences in amplitude of light, i.e., light and dark areas, which can be observed. As light rays pass through areas within the tissue of different optical path (refractive index and geometric path length) they may be retarded in phase by up to 14 but will remain unchanged in amplitude. Since the eye cannot discern phase differences, a mechanism for transforming phase changes into amplitude changes is required. Differential Interference Contrast Microscope: takes advantage of differences in the light refraction by different parts of living cells and transparent specimens and allows them to become visible during microscopic evaluation. Differential interference contrast produces contrast by visually displaying the refractive index gradients of different areas of a specimen. Parts: Linear Polarizer, Nomarski Prism, Objective Prism, Analyzer Fluorescent microscope is often used to image specific features of small specimens such as microbes. It is also used to visually enhance 3-D features at small scales. This can be accomplished by attaching fluorescent tags to anti-bodies that in turn attach to targeted features, or by staining in a less specific manner (see basic cell staining). When the reflected light and background fluorescence is filtered in this type of microscopy the targeted parts of a given sample can be imaged. This gives an investigator the ability to visualize desired organelles or unique surface features of a sample of interest. Confocal fluorescent microscopy is most often used to accentuate the 3-D nature of samples. This is achieved by using powerful light sources, such as lasers, that can be focused to a pinpoint. This focusing is done repeatedly throughout one level of a specimen after another. Most often an image reconstruction program pieces the multi level image data together into a 3D reconstruction of the targeted sample. Confocal Laser scanning microscope: Unlike compound and stereo microscopes, these devices are reserved for research organizations. They are able to scan a sample also in depth. A computer is then able to assemble the data to make a 3D image. Scanning Probe Microscopes: It is possible to visualize individual atoms with these microscopes. The image of the atom is computer-generated, however. A small tip measures the surface structure of the sample by rastering over the surface. If an atom projects out of the surface, then a higher electrical current will flow through the tip. The amount of current

is proportional to the height of the structure. A computer will then assemble the position data of the tip and the current to generate an image. The scanning tunneling microscopes use a piezo-electrically charged wire, a very small space between the charged wire and the surface and the specimen to produce enhanced images of the specimen. The charged wire forces energy across the small space and onto the specimen where the current meets with the specimens surface and decays. This decay is measured and a high-resolution image is produced from the information collected. Tunneling microscopy allows imaging at the atomic level to be produced plus different types of information can be obtained by altering the environment that the specimen is observed in such as a gaseous environment, vacuum, or a liquid environment. Atomic force microscopy uses a cantilever with a sharp probe that scans the surface of the specimen allowing for a resolution that you can measure in fractions of a nanometer; in other words "feeling" the surface of an object in order to produce a visual image. The flexibility of these types of microscopes are allowing for additional specialized instruments including the near field scanning optical microscope that utilizes optical fibers to stimulate specimens.