OREAS 131b - 131 b
We have a utility called the FoV calculator which is available on this site, access it by following the procedure below:
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In this lab, parts of the microscope will be reviewed. Students will learn the proper use and care of the microscope and observe samples from pond water.
A microscope is an instrument that magnifies an object so that it may be seen by the observer. Because cells are usually too small to see with the naked eye, a microscope is an essential tool in the field of biology. In addition to magnification, microscopes also provide resolution, which is the ability to distinguish two nearby objects as separate. A combination of magnification and resolution is necessary to clearly view specimens under the microscope. The light microscope bends a beam of light at the specimen using a series of lenses to provide a clear image of the specimen to the observer.
2. Always start with the stage as low as possible and using scanning objective (4x). Odds are, you will be able to see something on this setting (sometimes it’s only a color). Use the coarse knob to focus: the image may be small at this magnification, but you won't be able to find it on the higher powers without this first step. Move the mechanical stage until your focused image is also centered.
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To know if a camera can measure a target at a certain distance some calculations can be done. We have a utility called the FoV calculator which is available on this site, access it by following the procedure below: Go to Download Datasheet and select a camera series Click the FoV Calc. hyperlink for a camera–lens combination. This displays the FoV calculator. To select a different camera–lens combination for a particular camera, select the new combination in the drop-down menu. The FoV calculator displays the following results: HFOV: horizontal field of view in meters and feet. VFOV: vertical field of view in meters and feet. Depth of field—far limit in meters and feet. Depth of field—near limit in meters and feet. IFOV: instantaneous field of view in millimeters and inches. Distance to target in meters and feet. Tips Each camera may have interchangeable lenses. If this is the case, you can choose a lens from the drop down menu. You can switch between metric or imperial units using the drop-down menu. Type in several distances separated by commas to calculate the FoV at all desired distances. Example for an A655sc 45° Go to camera datasheets and select the product: Select the 45° lens from the dropdown menu: Type in the desired distance to calculate the FoV or use the listed default values.
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1. Place the letter “e” slide onto the mechanical stage. Be sure to note the orientation of the letter “e” as it appears to your naked eye.
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4. Switch to low power (10x). This may be sufficient to view your chosen organism. Try to note how it moves and do your best to draw it as you see it, unless you need more magnification.
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This portion of the procedure is another practice to demonstrate depth perception. Many new microscope users find it difficult to conceive that the specimen on the slide is in three dimensions. As the stage is moved up and down, different threads will be in focus.
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3. Use the SCANNING (4x) objective to focus, then move the mechanical stage around to scan the slide for live microorganisms. You are looking for tiny swimming beings- they may look green or clear and might be very small. Choose one to focus on and center it in your visual field.
5. Once you have centered and focused the image, switch to high power (40x) and refocus. Note movements and draw the organism as you see it.
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3. Once you've focused using the scanning objective, switch to the low power objective (10x). Use the coarse knob to refocus and move the mechanical stage to re-center your image. Again, if you haven't focused on this level, you will not be able to move to the next level.
In addition to the objective lenses, the ocular lens (eyepiece) has a magnification. The total magnification is determined by multiplying the magnification of the ocular and objective lenses.
Your microscope has 4 objective lenses: Scanning (4x), Low (10x), High (40x), and Oil Immersion (100x). In this lab, you will not use the oil immersion lens; it is for viewing microorganisms and requires technical instructions not covered in this procedure.
A large part of the learning process of microscopy is getting used to the orientation of images viewed through the oculars as opposed to with the naked eye. A common mistake is moving the mechanical stage the wrong way to find the specimen. This procedure is merely practice designed to make new users more comfortable with using the microscope.
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1. Using the transfer pipette, transfer a drop of pond water onto a microscope slide. The best specimens usually come from the bottom and probably will contain chunks of algae or other debris that you can see with your naked eye.
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2. Use the SCANNING (4x) objective and course focus adjustment to focus, then move the mechanical stage around to find the letter “e”. Note the orientation when viewed through the oculars.
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2. Use the SCANNING (4x) objective and course focus adjustment to focus, then move the mechanical stage around to find the threads.
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