BIOL 3053 Lab 2 Assignment (Winter 2022)

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Memorial University of Newfoundland *

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Biology

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Dec 6, 2023

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Memorial University of Newfoundland Department of Biology Biology 3053 Lab 2 Assignment Use of Microscopes and Smear Preparation Winter 2022 Name: Julie Hollahan Student #: 201311107 Exercise 3: Use of the Microscope 1. Draw 3-5 yeast cells from each magnification. Include any interesting structural changes evident at the three magnifications. medium dry high dry oil immersion Medium Dry High Dry Oil Immersion Ocular Lens Magnification 10X 10X 10X Objective Lens Magnification 10X 40X 100X Total Magnification 100X 400X 1000X (1.5 marks) 2. Examination of prepared bacteria slides. Examine with the oil-immersion objective and draw 3-5 cells of each morphological form. 1

Spherical Rod Spiral (Coccus/Cocci) (Bacillus/Bacilli) (Spirillum/Spirilla) 3. Describe what happens to the following properties of the microscope: (1.5 marks) (a) to the diameter of field of view when one moves from low to high dry power. The diameter would go from a large field of view to a small field, approximately 4500 um to 450um. (b) to the size of the image when one moves from medium to low power. The size of the image would decrease, going from a large to small size. (c) to the depth of field when one moves from low to oil immersion lens. The depth of field would decrease, going from a larger depth to the smallest depth, approximately 65um to 0.5um. 4. Give two reasons why we start our examination of any slide with the low power rather than oil immersion lens. (0.5 mark) 1) once oil is applied to the slide, you cannot change lenses and go back. 2) To be able to properly focus the object in the field of view. 5. Why is oil added to the surface of the slide before the oil immersion lens is clicked into place? (1.0 mark) The oil immersion lens would be in the way of properly applying the oil. 6. Besides reducing the amount of light, what other effect does closing the iris diaphragm have on the image? (0.5 mark) Closing the iris diaphragm also affects the image by changing the amount of contrast the object will display, If the iris diaphragm is closed or reduced, this reduces the amount of light getting in, which increases the contrast of the object. 7. Why is centering the object being viewed important before going to higher magnification? 2

(0.5 mark) This is important so that you do not lose the specimen in your field of view. Exercise 4: Smear Preparation 1. Is it advantageous to prepare a thin smear, such that only a few of cells are available for examination? Explain. (1.0 mark) Yes. The cell on the slide has a better chance of being stained. A thick smear will have cells stacked on top of each other, therefore reducing the staining process. A monolayer would be most appropriate. 2. Why should you allow a smear to air dry completely before heat-fixing? (1.0 mark) This reduces the likelihood of the specimen being splashed on your person or your workplace. 3. What occurs during the process of heat-fixing? (1.0 mark) Adherence to the slide for viewing, as well coagulation of the cytoplasm. 4. What are the three characteristics of a good smear? (1.5 marks) 1. A think layer, monolayer 2. Allowing full air to dry prior to heat fixing, reducing the likelihood of cell distortion occurring. 3. Ensuring proper heat fixing as the smear must undergo multiple washes. 3

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