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1 The Microscope sffiffi o. $et. a5 Name Partner Date The microscope, developed more than three centuries ago, is the basic tool of the biologist. A microscope enables biologists io investigate living things and objects that are too small to be seen with the unaided eye. The microscope is able to magnifr these tiny specimens by means of lenses located in the eyepiece and objectives. T[e [gtrt microscope is also capable of revealing fine detail. This ability to reveal fine aitiit is known as resolving power. The type of microscope that you will be using throughout your study of biology - is the compound light microscope. Specimens ihat are viewed under the microscope are mounted on one of two types of glass slides. Prepared slides are slides that are for permanent use. \ilet-mount slides are for temporary use. Most of the slides that you will use in biology will be wet mounts. In ybur study ofbiology, it will be necessary for youto estimate the length and width of some olfg,ur specimens. To measure objecti under the microscope, a unit called the micrometer (pm), rygetimes called tfre micton (p), is used. One micrometer equals millimeter; one millimeter equals 1000 micrometers. Give the purpose of the microscope: Define resolving power: Name the type of microscope used in high school biology plasses: Contrast the use of prepared and wet-mount slides: Name the preferred unit of measurement for microscope work: Tell how many micrometers are in a millimeter: also called the Hour Give the syrnbol for the micrometer: Stage opening permits light from light source to pass up through body tub Oiaphngm regulates amount ol l.lght passing through speclmen Lamp or mirtor directs lighl through diaphragm and stage openrng

2 Carrying the Microscope: Take the microscope from the storage arca. Carry the microscope with one hand under the base and the other hand grasping the arm. Place the microscope on the laboratory table. The microscope should be about 10 cm from the edge of the table. Uncoil and plw the cord into an outlet at your lab station. The on(l)-otr(0) switch is located on the left side ofthe base ofthe scope. Parts of the Microscope: Look at the drawings on the previous page that is most like your microscope. Identifr the parts and functions of your microscope. Cleaning the Microscope: Carefully clean the eyepiece and objectives lenses with lens paper. Locate the nosepiece and gently turn it so that the low-power scanning objective is in line with the body tube. The nosepiece will click into place when the objective is in the proper position. Field of View: Keeping both eyes opeo look through the eyepiece. You will see a circle of light. This is called the field of view. To make the circle of light as bright as possible, you may have to adjust the diaphragm. Your Dominant Eye: If you are observing through a single lens (monocular) scope, learn to see with your dominant eye while training the other eye to relax and not concentrate on anything. This technique is useful and is easily learned with practice. It will come in handy when you draw your specimens as you can use your weaker eye to help you see your drawing while observing with the dominant eye. People normally have a stronger (dominant) eye (the one that is most used). It is often the one used for microscopic observation, unless you are using a binocular stereo microscope. Then, you would use both eyes to observe your object or specimen. Right or Left? Which is your dominant eye? In order to determine which eye is dominant (stronger), there is a simple test. Use your hands to form a circle at arm's length. Look across the roonl through the circle, both eyes open (relax), at some object. Slowly bring the circle toward your face. Your "handcircle" will probably go to one eye or the other-this is your dominant eye. Use this eye for viewing. Describe your hand-positionwhen carrying the microscope: Give the rulme for the circle of light you see wlren looking through the eyepiece: Name the microscope part used to adjust the amount of light in the field of view: Contrast a monocular and binocular scope: Which eye is your dominant eye? There are three objective lenses on your microscope. The shortest lens is called the scanning lens-it has a red ring around it. The low-power objective is the medium-length lens and has a yellow ring around it. The high-power objective is the longest lens and has a blue ring around it. The lens you are using is the one that is "clicked" into place above the stage opening. You could also say it is the one below the body tube. A4iust your micrqscope so the scanning lgns is in place for viewine a specimen. z

3 Preparing and Observing a \ilet Mount Obtain a glass slide and cover slip. Wipe both sides of the slide with a cloth to remove dust, etc. The cleaned slide and cover slip should be handled by their edges. Cut out a small piece of newspaper that contains the letter "e." This should be a standard newsprint letter- NOT a headline-size letter-avoid dark backgrounds on either side of the newsprint. Place the letter "e" in the center ofthe slide as it would appear in the newspaper-face-up. rieht side up. Using a pipette, place a drop of tap water on top ofthe "e." IJse a probe to hold the "e" in place while you add the drop of water, if necessary. Hold a cover slip at about a45o ang,le over the drop of water. Gently lower the cover slip onto the slide. If air bubbles appear, gently tap the cover slip with the back end of the probe. Dissecting needle Place the wet mount of the letter "e" on the stage of the microscope rryith the letter facing yor+ as you would read it in print. Adjust the slide so that the letter is above the opening of the stage. Look at the slide at eye level. Observe the space between the slide and the scanning objective. The scanning objective should be in line with the body tube (over the stage opening). Slowly turn the coarse-adjustment knob, raising the stage to its highest position. You are now ready to view your specimen. Look through the eyepiece and SLOWLY turn the coarseadjustment knob until the letter "e" comes into focus. Inthe circle below (which represents a field of view), sketch what you see. CAUTIONz Never raise the stage while looking through the eltepiece; you may hit and danrage the slide or ohjective lens. Look to the side, raise the stage to its nruxintum height, then focus u,hile the stage is movirtg AWAY fi'om the objective. Calculate the magnification (eyepiece x objective). Show your math in the space below. Leave your slide on the stage and have your instructor veriff your view. Your instructor will initial the blank below. Completed

4 Make a second slide of a letter o'e" but this time from print provided by your instructor. Once again, check the orientation of the letter (it should be in the same position as it appears in print). Rotate the nosepiece to the actual low-power lens. This is the mid-length lens (with a yellow circle) and is marked with a magnification of 10X. Following proper technique, locate and focus on the letter. Calculate the magnification below (show your math): Sketch what you observe below. Completed Paradoxical Movement: The microscopexhibits an optical phenomenon called paradoxical movement. Move the slide to the right...which way does it "appear" to move in the field of view? Move it to the left....which way does it o'appear" to move? Move the slide away from you, then toward you. Which way does it "appear" to move each time? when an optical instrument exhibits paradoxical movement? What does it mean Practice centering an object in the field of view. For example, an object is on the edge of the field of view and you want to center it before you ask your partner to take a look. Determine which way you would move the slide to center the specimen. Below are some sample problems. Use an arrow (or two arrows) to show which way you would move the slide to center the object: Y

5 To observe a specimen at high-power magnifrcation, turn the nosepiece until the high-power objective clicks into place. (The high-power objective is the longest object, has a blue ring, and is marked 40X.) You microscope is parfocal. This allows you to focus the scope at low power, switch to high power, and, with only minor adjustment of the fine-adjustment knob, see the object at high power. For that reason, here is another general rule of microscopy: To view an object at high power, always begin by focusing on it at low power. Then, switch to high power and use the fine-adjustment knob to bring the object into view. Observe the letter at high power. Draw what you observe Calculate the magnification below (show your math): Completed Measuring an Object Under the Microscope Turn the nosepiece so the scanning lens (4x, 40x total) is in place. Place a millimeter scale of a transparent plastic ruler over the center of the stage opening in the microscope. Use the scanning objective to locate the millimeter lines of the ruler. Place these lines in the middle of the field of view and use the coarse-adjustment knob to bring them in to focus. The distance between two lines on the ruler represents I mm. While looking through the eyepiece, move the ruler so that one of the millimeter lines is just touching the left side of the field of view. You ruler will look like the diagram but you will be able to see more millimeter lines since you are working at a lower magnification. Field of view To determine the diameter of the field of view for the scanning lens, count the number if millimeter lines (actually spaces) that are visible. You will need to estimate the diameter to the nearest tenth of a millimeter. 2 millimeters Since the micrometer (pm) is the preferred unit of measurement for use with the microscope, convert your measurement in millimeters to micrometers by moving the decimal three places to the right (effectively multiplying by 1000). Scanning lens (4x, 40x total) field of view: mm or

6 Next rotate the nosepiece to the low-power objective (10x, 100 x total) and determine the field of view. Count the number of millimeter lines that are visible. Since the millimeter marks themselves are now rather large, you will need to move the edge of one of the marks to the left side of the field of view and count from the left edge of one mark to the left edge of the next. Don't forget to estimate the nearest 0.1 mm. Low-power objective (10x, 100x total) field of view: mm or pm Because under high power (40x, 400x total) the thickness of one of the millimeter lines takes up practically the entire field of view, it is difficult to estimate the diameter of the field veiw under high power magnification. The diameter under high power can be calculated on paper, however. Here are the steps: l. Divide the magnification of the high-power objective by the magnification of the low power objective. Show your math in the space below: 2. Then divide the diameter of the low-power field size of view in micrometers by the answer to the step one above. Again, show your math: High-power objective (40x,400 total field of view: mm or pm Sample problem: Here is what you see in your scope at 100x (using the 10x objective): For low power (10x, l00x total): Field of view Edge of ruler For high power (40x, 400x total): Millimeter lines 2 millimeters ("

7 Record your group's field sizes for the scanning (4x, 40x total), low-power (10x, l00x total), and calculated high-power (40x, 400x total) on the data table in class. The class will adopt a standard set of field sizes from class data. Do not write anything in the section below until all the class data has been recorded. Class standards: Prepare a wet-mount of a human hair. Observe and draw the hair at scanning, low, and high powers. Low-power x High-power To calculate the width of a human hair, estimate how many could be placed side-by-side across the field of view. Divide the size of the field of view by this number. Show your math for each problem in the space below: 7

8 Practice problems: /t Human Hair l00x Human Hair 400x Prepare a slide of two crossed threads of different colors. View the threads at low power and draw what you see in the space below. You drawings should include color. Estimate the width of the threads. Crossed Threads Width of a thread pm \lij U\ c,*1"^\q{, ons : <. qrhpl"[" o\ v

9 Make a temporary wet mount of material from your'opond inajar" and view it using the microscope. Attempt to choose one microorganism and draw it in the space below. Pond in a Jar Width or length _ \^ri A+h c.o.tc\^lslions : pm Continue your work with the pond water by preparing additional slide(s) and locating at least two other microorganisms. Draw them in the spaces below and use the field guide to identi$z them. x Width or length pm Width or length pm Cq[o.rt\.^li snr.. C"o.l.-,rl q{'on; } 1

10 Label the parts of the microscope in the figure below: Looking through the microscope, in what direction does the letter "e" appear to move when you moved the slide to the rieht? to the left? away from you? toward vou? What is the above phenomenon called? Calculate the total magnification of your microscope for the following objectives. (Show your math) Scanning Low-power High-power What happens to the focus of the letter'oe" as you change from low-power to high-power magnification? When the above occurs, your microscope is said to be How many times is the magnification increased when you change from low-power to high-power masnification? What happens to the size of the field of view when you change from low-power magnification to high-power maenification? How many micrometers are in a millimeter? Give the alternate name for the micrometer: Give the unit for the micrometer: Give its unit: )a

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