7-Report Microscopes_2023

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Colorado State University, Fort Collins *

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Jan 9, 2024

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LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ The Microscope I: Proper Use 60 pts total plus Notebook Conclusion (4 pts) A. Parts of the Compound Microscope and their Function 1. Label the microscope parts in the corresponding blank boxes. (10 points) 2. What is the function of the objective lens? (1 point)

LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ 3. Magnification: Indicate in the table below the total magnification of an object when viewed with each of the following objectives. HINT: Use the equation: Mag(total) = Mag(objective) x Mag(ocular) (4 points) Objective Lens Magnification 4X (Scanning Lens) 10X 20X 40X 4. What is resolving power and how does that relate to magnification? (1 point) 5. Why should we view our samples starting with the lowest magnification? Why is that lens called the scanning lens? (2 points) 6. Be able to explain how to adjust your microscope for Kohler illumination prior to calling a TA over to sign off on your report. Start with the typical plant leaf slide. Record interpupillary distance adjustment (IPD) below. Ask a TA for help, if needed. (1 point) IPD Adjustment # ________________ Koehler Illumination adjustment: Be able to explain the process of the Kohler illumination adjustment from start to finish. What is the function of the condenser? Use the typical plant leaf slide. (1 point)

LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ Signed TA ___________________________________________________________ B. Exercises Stressing Microscope Technique: 1. The Letter “e”: a. What orientation is the letter now? Upside down or right side up? (1 point) b. Move the slide (or stage) towards you while looking through the microscope. Does the letter move away from you or toward you? (1 point) c. How does the image move when the slide is moved to the right or left? (1 point) 2. Annular Stop Turret: Start with a focused sample of transparent cells (unstained fibroblasts) under the phase contrast setting. Before proceeding, check with the TA or Instructor to ensure that the fibroblast cells are present. Record in the table below whether you see the transparent image or not. Describe the image resolution at each of the illuminations and magnifications below as you change the annular stop turret. (6 points) Annular Turret Adjustment Ability to See (Yes or No) Definition (High, Medium, or Low) Phase Contrast 10X Phase Contrast 20X Phase Contrast 40X Bright Field 10X

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LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ Bright Field 40X Dark Field 10X, 20X, 40X 3. Which illumination is best for viewing transparent specimen? Support your answer. (2 points) 4. This next set of exercises relies on 2 types of rulers. One is a regular student inch/centimeter ruler. The other one is a special ruler on a slide, called the stage micrometer. Determination of Size and Brightness of Field of View (FOV): a. Student ruler method: Using the provided student ruler measure the diameter of FOV at 4X objective and record it here. (1 point) b. Calculate the FOV for 4X, 10X and 40X objectives. Show your work. (Note: FOV low * Mag low = FOV hi * Mag hi ) (3 points) 4X FOV: 10X FOV: 40X FOV: c. Can you measure the field of view at a higher magnification with a regular student ruler? Explain. (2 points) d. Stage/Ocular Micrometer Method: Calibrate the ocular micrometer with the stage micrometer. Show your calculations for each of the objectives in the table below. (6 points) One Ocular Space (mm or um) = X (mm or um)/ Y Y= number of ocular spaces, X= mm or um Objective Power 4X 10X 20X 40X X (mm or um)

LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ Y (# ocular spaces) One Ocular Space (mm or um) Note: Save these calibrations in your notebook. You will need them to measure cell sizes next week in Microscopes Lab II. e. Fill in the table below with your ocular space calibration in um for each of the specified objectives. Use your calibrated ocular ruler to measure the diameter of the field of view (FOV). Calculate the total area of FOV for each of the three objectives. (8 points) Note: Be sure to use your calibrated ocular micrometer to measure the diameter of the FOV. DO NOT use the FOV figures that you calculated using the student ruler in the question 4a above. Objective Power Size of one ocular space (um) FOV Diameter (mm) FOV Area (mm 2 ) 1 Estimated % of Light Allowed Through 2 4X 10X 20X 40X f. Does the field of view increase or decrease with increasing magnification? (2 points) g. Calculate the difference in areas estimated for the 4X objective compared to the 40X objective. (2 points)

LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ 1 Area of circle =  r 2 2 Assume the light intensity that you see at 4X is 100% and estimate how much more or less light intensity you see at higher magnifications. h. What happens to the light intensity (brightness of the field) as the objective power increase? (1 point) i. Compare and contrast your results in the two methods of measuring FOV (Ruler Method vs. Stage/Ocular Micrometer Method). Which one should be more accurate? Explain. (2 points) 5. Depth of Field/Focus: Which objective, high power or low power, provides the greatest depth of field with all threads in focus? Explain why. (2 points) Notebook Conclusion (4pts)

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LIFE 212 Laboratory Report 7 Name: ___________________________ Due:_____________________ Section:___________________________ The conclusion should be a concisely written summary of the experimental results using technical language. Describe each objective from your Introduction individually and frame in context with the experimental results. Vague and/or incomplete conclusions will not receive any credit.

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