Microscopy and Cellular Measurements: Lab 1 Guide

Biol 1010 - Lab # 1 — Fall 2023 INTRODUCTION TO MICROSCOPY AND CELLULAR MEASUREMENTS Laboratory assignment Name: ID #: TA's name: Lab CRN #: Please bring your laptop and ear pieces to the lab. You will not be able to attend the lab otherwise. You do not need to bring a printout of your Intro/Protocol file (only for this lab!) but you need to bring a printout of your assignment file with your pre-lab assignment question completed. Pre-lab assighment questions: - Complete the use/function of Table 1 (p 2 below - Watch the microscopy video (on Canvas). Fill in Table 2 (p 3 and answer all the questions on page 4 of this assignment. - You won't be able to attend the lab if this is not done. In this lab you will work in pairs but you will both be required to know how to perform the following tasks individually. Remember that you must answer all questions on you own. Getting answers from other students or giving your completed work to someone else will be considered plagiarism and will be penalized. l1|Page

Exercise 1: Safety in the lab Table 1: Location and uses of the safety features found in the lab. Complete the use/function as part of the pre-laboratory assignment Sticker colour * Safety feature Uses / Functions Eye wash station Shower First aid Kit Spill Kit Fume hood Fire extinguisher Emergency exit Glove waste Emergency contact numbers * Please make sure you also sign the sheet located at each safety station. 2|Page

Exercise 2a: Parts of a compound microscope =~ 10 11 Table 2: Using the picture above, indicate the number and the name of the part corresponding to the description mentioned in the first column. Complete before the lab as part of the pre-laboratory assignment Description Number | Name of the part Move specimen side to side or up and down Controls the amount of light leaving the source Magnify the sample with different magnifications Focuses the light onto the specimen Hold the specimen in place Turn the power of the microscope Magnifying lenses, also called eye pieces Where you place the slide Only used with low magnification objective lenses Regulates the intensity of light provided to your sample Allows for sharp focus of the image 3|Page

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Exercise 2b- Microscope setup Pre-lab assignment: Answer the following questions while watching the Microscopy video (on Canvas). Write your answers in the space provided. What should you do before placing a slide on the stage? (two steps) What is the fastest way to focus your microscope? What is Kohler illumination used for? (give one) Briefly explain how to set up your microscope for Kohler illumination. Indicate the microscope parts used in your answer. (2 steps) You have focussed your microscope with the 10X objective; what should you NOT do after switching to 40X? Which steps should you take BEFORE removing a slide from the stage? (two steps) 4|Page

Setting up your microscope: The following individual exercises must be performed during the lab. Make sure you complete this activity before leaving the lab. Review the information posted in the Introduction section and the Microscopy video (on Canvas). Practice setting up your microscope. Practice the technique, your TA will assess of your ability to: - Focussing at 10X - Set up Kohler illumination - Focussing at 40X Your TA will do an individual assessment, testing your proficiency to set up the microscope. You will have 4 min to do the requested set-up without any help. The exercise is out of 8 marks. If you obtain less than 6/8 on your first attempt, you will be required to study the technique again and repeat the testing one more time. Please note that you will lose 2 points out of your final mark if you have to be retested for the technique. 15t attempt 2" attempt Focus a prepared slide with the 4X or 10X objective (3) Set your microscope for Kohler illumination (4) Focus a prepared slide with the 40X objective (1) Penalty NA 2 Final mark: 5|Page

Exercise 3- Magnification and Resolution Table 3: Calculation of total magnification and theoretical resolution. Objective Ocular lens Total Numerical Theoretical lens maghnification | magnification | aperture | resolution (R) maghnification at A= 531 nm (in pm) 4X 10X 40X 100X 1.25 Calculate of the total magnification and the resolution (R) for all the lenses. Convert the R values calculated from nm to pm and round the numbers to one digit after the decimal point. Show the formula used and ALL your calculations below and fill in your answers in Table 1. Remember to include the units. 6|Page

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Calculate the limit of resolution when using a 100X lens with and without oil What is the effective numerical aperture of your microscope when using the 100X lens without oil and when using the 100X lens with the oil. Briefly explain how you came up with these values. Note: See page 17 of your protocol, Numerical aperture without oil: Explanation: Numerical aperture with oil: Explanation: - Calculate the resolution values for the 100X lens with and without oil at A =398 nm. Show all your calculations below and write the final values in ym with one digit after the decimal point and using scientific notation. - Using the resolution values you just calculated, is it best to use oil or no oil when using the 100X lens? Briefly explain. 7|Page

From the calculations performed above, can you see objects that are 0.00022 mm apart as being distinct when using the 100X lens with oil? (Show your calculations and briefly explain). Calculations: Exercise 4: Table 4: Drawings of a plant and an animal cell at 40X Elodea Parascaris Uterus 8|Page

- Determine the actual size of a plant and animal cells using the ocular micrometer (write the formula used and show your calculations). - Determine the magnification of your drawings using the size of your drawing (measured with a ruler) and the actual size of the specimen determined above (write the formula used and show your calculations). - For each of your drawings, determine the actual size that corresponds to 1 cm of your drawing (write the formula used and show your calculation). Add the scale bar to lower right corner of your drawings and write on top of it the actual sizes that corresponds to 1 cm. 9|Page

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Answer the following questions: - Name three similarities between plant and animal cells - Name three differences between plant and animal cells - Which similarities or differences could you see during the observation of your plant and animal cells 10|Page

bio 1010 lab 1 assignment

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