Name, Number. EXPLORING THE COMPOUND MICROSCOPE Introduction: ‘Microscopes are very important scientific tools that allow people to observe small things they normally couldn't see with the naked eye. In our laboratory, we use compound light microscopes. They are called compound light microscopes because they contain two lenses—an objective lens and an eye piece—that help to magnify (enlarge) an image, The microscopes used in our laboratory have at least three different objective lenses, each with a different magnification, Using the different lenses we can magnify the image a little (low power) or a lot (high power). Working with the compound microscope, you may use specimens that have been prepared in one of two ways. A prepared slide is made to be permanent and can be purchased from a catalog. A wet-mount slide is made for temporary use and can be made. and used during the lab period, Often, the specimen must be stained using special dyes. This enables the observer to easily see parts of the specimen that would be invisible without the dyes. Pre-Lab Questions: What does the word 'magnify’ mean? What two lenses help to magnify an image on a compound light microscope? What are the two types of slides we can view? Why do we use stains in the laboratory? Objectives: 1. Learn the parts and operation of a compound light microscope 2. Learn to prepare and observe a wet mount slide 3. explore the ideas of resolution and depth of field using the microscope Part I. Calculating Total Magnification The compound light microscope has at least three objective lenses, Each lens has a different magnifying power—low power and high power is what we'll be using. You can see how many times the lens is magnifying the image by looking at the number located on the lens. For example 4X means it magnifies the image 4 times its normal size. The eyepiece also has a number on it indicating how much it magnifies the imageThe total magnification of the microscope is easy to calculate. Use the formula below to calculate total magnifying power: Magnifying power of eyepiece X Magnifying power of objective lens =) 1. What is the magnifying power of your eyepiece? 2. What is the magnifying power of your low power objective lens? 3. What is the magnifying power of your high power lens? Using the formula from the previous page, calculate the total magnification of your microscope under LOW POWER in the space below: DE eee eee eee Eyepiece Low power Total magnification objectivelens —_- magnification magnification Now, calculate the total magnification of your microscope under HIGH POWER: Eyepiece igh power Total magnification objective lens magnification magnification Part IL. Preparing and Examining a Wet Mount Slide Your teacher will model how to prepare a wet mount slide of a letter ‘e’ (refer to the diagram below if you are still unsure). Pioce of torating Cut out fetter from newspaper and place letter ‘e’ on the slide Put 1-2 drops of Place cover slip on water on top of 'e’ —_top of ‘e’ at an angle Now you hovel to avoid air bubbles Wet mountA. Focusing the image under LOW POWER MAGNIFICATION: Once you have prepared your slide, place it on the stage of the microscope and secure it with the stage clips, 2. Turn your microscope to the lowest power objective lens and turn the coarse adjustment knob until the objective lens is as far up and away from the stage as possible. 3. Rotate the diaphragm until it gives the maximum amount of light. 4. Look into the eyepiece and slowly turn the coarse adjustment knob until the letter ‘e’ comes into fecus. Re-center your'e’ as needed by moving the slide on the stage, 5. Use the fine adjustment to sharpen the image if necessary. Draw the letter 'e' as you see it through the microscope Describe what the microscope has done to the letter ‘e’. eee Ee eS eR EET —— Look through the eyepiece and move your slide to the RIGHT. What direction in the slide moving when you look ‘Through the microscope? Look through the eyepiece and move your slide to the LEFT. What direction in the slide moving when you look through the microscope? Now move the slide UP and DOWN, What happens? eee eters eee at ae teaser eee B. Focusing the ‘e' under HIGH POWER 1. Make sure the 'e’is in the middle of your field of view. The field of view is how much of the object you can see under the microscope under various magnifications, turn to high power magnification focus the image by using the FINE ADJUSTMENT KNOB only! You have already focused the image. It will just need a little fine tuning, 4, Adjust the diaphragm if needed wn Draw what the 'e' looks like under HIGH POWER in the circle to the right. Are you Seeing more of the letter'e' or less of the letter ‘e? Based on your answer to the last question, is the field of view larger under high power or larger under low power? Did the image get darker or lighter when you switched from \ W power to high power?Part III. Resolution The resolving power or resolution of a microscope is its ability to show 2 points that are very close together as separate images. c Follow the directions below to observe the microscope resolving power: Matnytone ue Cut out a tem square. piece of colored f . Saeed ut 1-2 drops of Place cover slipon Tee laa water ontop of top of colored piece Now you a a nage paper ‘of newspaper at an Wet mount a id angle to avoid air bubbles After you prepare your wet mount, focus the colored paper under low power magnification. Refer back to the directions on page two if you are unsure how to focus ( the image. Now turn to high power and focus the image as directed before. Describe what the paper looks like under high power compared to the naked eye. In your response describe the colors you see and any patterns you see. Part IV. Depth of Field ‘The microscope can only focus on one depth (or plane) at a time, Because the object you are viewing is 3-D, it can only focus on part of the object ata time. Follow the directions below to observe the microscope's varying depths of field: 1. Pull out two pieces of your hair and arrange them in an'x’ shape on your slide. 2. Prepare awet mount slide as directed before and focus the hairs under LOW POWER, focusing on the point where the hairs cross. 3. Now focus the image under HIGH POWER as directed before. Is the entire image in focus? Look into the eyepiece and turn the fine adjustment knob back and forth. lu What happens to the image when you do this?