Bx SAB REPORT NO. | Ona ___ MALIC, ANZEL ANGELO @), a ; p MLB 2 de1. Arm - Main support for microscope components. 2. Base - Housing and platform of the instrument to which the arm is attached in addition it usually contains an illumination system for the microscope. 3. Diopter adjustment - Adjustable eyepiece diopter permits focusing adjustment of image for any difference in vision between users eyes. 4. Disc Diaphragm - Disc located below stage with holes of various apertures, designed to help achieve optimum resolution of the objective lens. Larger apertures used for higher magnifications, and smaller apertures used for lower magnifications, 5. Eyepiece (ocular lens) - Lens closest to the eye magnifies the primary image formed by the objective lens. 6. Eyepiece tube - This is the component that holds the eyepieces in place. Elementary, student and high school models have set screws in the eyepiece tube used to lock eyepieces in place. 7. Filter - Daylight blue filter designed to make incandescent illumination appear white. 8, Filter holder - Attached to bottom of iris diaphragm that swings out allowing user to insert filter of choice. Ifa built in neutral filter is provided it should be removed from the optical path when using 40x and 100x objectives. 9. Head - Upper portion of the microscope which contain prisms and eyepiece tube or tubes. 10. Iris diaphragm - Iris Diaphragm, opening and closing of iris is controlled by lever. It is designed to help achieve optimum resolution of the objective lens. Larger apertures used for higher magnifications, and smaller apertures used for lower magnifications, 11. Mechanical stage - Permits precise, mechanical manipulation of the specimen slide. 12, Nosepiece (revolving turret) - Designed to hold objective lenses permitting changes of magnification by rotating different powered objective lenses into optical path. Forward facing position used on elementary and high school models. Reverse facing nosepiece position used on’ more advanced models permits easier access to stage when posi 13. Stages clips - Two locked-on clips hold specimen slide in place on stage. 14, Stage - Platform of the microscope where the specimen slide is placed.Bera) perare (9A) © Mathematical formula devised by East Abbey the direct comparison of objective lens to resolving power. 2. Course focusing knobs - Large knobs located on each side of arm, raise or lower stage to bring specimen image into focus. 3, Fine focusing knobs - Smaller knobs, located close to the coarse focusing knobs, permit more: precise adjustment of the image. Magnifications: 1. Abbe condenser - ‘The 1.25 N. A. Abbe condenser lens positioned under the center of the stage is requited when using 100x objective lenses. In addition the Abbe condenser is focusable by one of two methods, a spiral mount condenser with lever to lower or raise the assembly or else a rack and pinion mount with a knob to provide movement in the up and down direction. 2. Condenser Lens (0.65) - Condenses light rays from sub stage illumination and fills the back element of objective lens to improve image resolution. A 0.65 condenser lens fixed in center of stage is provided on microscopes with objectives up to 40x (400 times magnification). 3. Objective lenses - Lens closest to the object being viewed, forms first image of the specimen. 4, Scanning Objective lens - A scanning objective lens provides the lowest magnification power of all objective lenses. 4x is a common magnification for scanning objectives and, when combined with the magnification power of a 10x eyepiece lens, a 4x scanning objective lens gives a total magnification of 40x. 5. Low power objective lens (10x) - The low power objective lens has more magnification power than the scanning objective lens, and it is one of the most helpful lenses when it comes to observing and analyzing glass slide samples. The total magnification of a low power objective lens combined with a 10x eyepiece lens is 100x magnification. 6.High power objective lens (40x) - The high-powered objective lens (also called “high dry" lens) is ideal for observing fine details within a specimen sample. The total magnification of a high~ power objective lens combined with a 10x eyepiece is equal to 400x magnification, giving you a very detailed picture of the specimen in your slide. 7.0il immersion lens - High power (100x) objective lens which requires a medium of immersion oil between the lens and the slide. Iluminations systems: 1.Rheostat - Variable potentiometer that adjusts the light intensity of the illuminator. 2, Illuminator - To provide even, high intensity light at the place of the field aperture, so that light can travel through the condenser to the specimen, 3. Transmitted illumination - Used to illuminate both compound and stereo microscopes providing illumination from below specimen. IL Enumerate the precautions in the proper handling and use of a compound microscope. ‘+ Always use two hands when moving the microscope. ‘+ Make sure the microscope is not plugged in before moving it. ‘+ Ifusing immersion oil, be sure to clean the immersion oil lens thoroughly after use. + Keeping your microscope clean (especially the optics) will also ensure that it lasts longer. ‘* When the microscope is not in use cover it with a dust cover or store it in a microscope case.ing of the following terms 1. Parfocal lenses ~ A parfocal lens system is a lens that maintains its focus despite changes in the lenses focal length or magnification. 2. Focus - A means of moving the specimen closer or further away from the objective lens to render a sharp image, on some microscopes, the stage moves and on others, the tube or head of the microscope moves. Rack and pinion focusing is the most popular and durable type of focusing mechanism. 3, Resolving power refractive index - The wavelength of light, refractive index, and angular aperture are important factors determining resolving power. Resolving power refers to a microscope's ability to distinguish two adjacent points as distinct and separate. 4, Magnification - Total magnification obtained with each objective lens is determined by multiplying the magnification of the eyepiece times the magnification of the objective. Keep in mind that as magnification increases, field of view (area of the specimen seen when looking through microscope) decreases. 5. Birefringence magnification - can be defined as the optical characters of a material having a refractive index which relies on the polarization and propagation direction of light. 6. Total magnification - In a compound microscope the total magnification is the product of the objective and ocular lenses (see figure below). The magnification of the ocular lenses on your scope is 10X. Objective lens X Ocular lens = Total magnification. 7. Monocular microscope - A monocular microscope is a microscope that only has one lens. 8, Binocular microscope - A binocular microscope is any optical microscope with two eyepieces to significantly ease viewing and cut down on eye strain. V. Convert the following units to Si: ‘centimeter (cm) 0.01 meter 1 millimeter (mm) 0.001 meter ‘1 micrometer (um) 1.0E-6 m meter * 1 millimicron (mu) I nanometer 1 Angstrom 0.1 nanometer VI. Give the differential features of each of the following types of microscope: 1. Phase contrast - A light microscopy technique used to enhance the contrast of images of transparent and colourless specimens. It enables visualization of cells and cell components that would be difficult to see using an ordinary light microscope. As phase contrast microscopy does not require cells to be Killed, fixed or stained, the technique enables living cells, usually in culture, to be visualised in their natural state. This means biological processes can be seen and recorded at high contrast and specimen detail can be observed. Fluorescence staining can be used in combination with phase contrast to further improve the visualisation of samples.2. Interference /Interference-contrast ~ An optical microscopy technique that uses interference between two white-light illumination beams or rays to generate an image with enhanced contrast. 3. Polarizing microscope - A contrast-enhancing technique that improves the quality of the image obtained with birefringent materials when compared to other techniques such as darkfield and brightfield illumination, differential interference contrast, phase contrast, Hoffman modulation contrast, and fluorescence. Polarized light microscopes have a high degree of sensitivity and can be utilized for both quantitative and qualitative studies targeted at a wide range of anisotropic specimens. 4. Darkfield microscope - Dark field microscopy is a technique that takes advantage of oblique illumination to enhance contrast in specimens that are not imaged well under normal illumination conditions. After the direct light has been blocked by an opaque stop in the condenser, light passing through the specimen from oblique angles is diffracted, refracted, and reflected into the microscope objective to form a bright image of the specimen superimposed on a dark background. This dark background provides a high degree of contrast and can make samples with difficult backgrounds stand out with relatively little effort. Fight: sileworm lana space and trachea. Ty peces of ragmented wood take on an unusual beautfl appearance ‘when lurinated under darks conditons witha ransmited ight mcvoscepeGe)ri ron microscope (TEM). Indicate the use of each: The two EM systems also differ in the way they are operated. SEMs usually use acceleration voltages up to 30 kV, while TEM users can set it in the range of 60 ~ 300kV. The magnifications that TEMs offer are also much higher compared to SEMs: TEM users can magnify their samples by more than 50 million times, while for the SEM this is limited up to 1-2 million times. Larger than TEMs, which users can only use to image a very small part of their sample. Similarly, the depths of field of SEM systems are much higher than in TEM systems. In addition, the way images are created are different in the two systems. In SEMs, samples are positioned at the bottom of the electron column and the scattered electrons (back-scattered or secondary) are captured by electron detectors. Photomultipliers are then used to convert this signal into a voltage signal, which is amplified and gives rise to the image on a PC screen, In a TEM microscope, the sample is located in the middle of the column. The transmitted electrons pass through and through a series of lenses below the sample (intermediate and projector lenses). An image is directly shown on a fluorescent screen or via a charge-coupled device (CCD) camera, onto a PC screen. it,Scanning Electron Microscopy (SEM) Transmission Electron Microscopy (TEM) Light Source SEM is based on scattered electrons, i. electrons emitted from the surface of a specimen. Itis the EM analog of a stereo light microscope. Electrons are used as “light source”, TEM is based on. transmitted electrons and operates on the same basic principles as the light microscope. Purpose SEM provides detailed images of the surfaces of cells. SEM focuses on the sample's surface and its composition, so SEM shows only the morphology of samples. Transmission electron microscope is used to view thin specimens (tissue sections, molecules, etc). TEM can show many characteristics of the sample, such as internal composition, morphology, crystallization, ete, Sample Preparation Sample is coated with a thin layer of heavy metal such as gold or palladium. ‘The sample in TEM has to be cut thinner (70-90 nm) because electrons cannot penetrate very far into materials. Magnification Image formation ‘The magnifying power of SEM is up to 50,000X. Secondary or backscattered electrons arising from the interaction of electron beam and metal-coated specimen are collected and the resulting image is displayed on a computer screen. ‘The magnifying power of TEM is up to 2 million times. ‘Transmitted electrons hit a fluorescent screen giving rise to a “shadow image” of the specimen with its different parts displayed in varied darkness according to their density. The image can be studied directly by the operator or photographed with a camera Current Applications To study topography and atomic composition of specimens, process control and also, for example, the surface distribution of immuno-labels. To image the interior of cells (in thin sections), the structure of protein molecules (contrasted by metal shadowing), the organization of molecules in viruses and cytoskeletal filaments (prepared by the negative staining technique).~ joining several photos taken through the microscope. The frequently used term “virtual” refers to. the examination of the specimens without direct contact to the object slide or the light ‘microscope. The term “high-resolution digital microscopy" is more precise, but not frequently used, Other terms that are frequently being used are: internet based microscopy, ePathology, eHistology, eHematology, eBiology or virtual microscope. These terms are to some extent used synonymously with the term virtual microscopy. Microscope slides are digitalized either manually or fully automatically with a 20x or 40x objective and saved in various formats, depending on the different service providers. IX. References: 1. https:/ /www.microscopeworld.com/p-3568-5-tips-to-properly-care-for-your- microscope.aspx 2. Microscope instruction manual - Fischer scientific. 3. https://www.studiobinder.com /blog/what-is-a-parfocal-lens-definition/ 4. https://www.sciencedirect.com /topics /immunology-and-microbiology /birefringence 5, www.scientifica.uk.com/learning-zone/a-guide-to-phase-contrast 6 4 8. 9. . https:/ /www.microscopyu.com /techniques/polarized-light /polarized-light-microscopy . https:/ /www.umassmed.edu/cemf/whatisem/ . https: / /microbeonline.com /difference-electron-microscopy-between-sem-tem/ . https:/ /www.thermofisher.com/ph/en/home/materials-science /learning- center/applications/sem-tem- difference html#:~:text=The%20main%20difference%20between%20SEM,sample)%20t0%20cre ate%20an%20image. 10. https://www.virtual-microscopy.net