Observation under the microscope
Cells cannot be seen with the naked eye: you need an instrument that magnifies them hundreds of times. The compound light microscope is the basic tool of any biology lab, and using it well does not come automatically: it has its steps and its tricks. Imagine Emma's science fair project at her middle school in Houston: she wants to see what a piece of onion is made of. Can she do it with just her eyes? Let's find out how to do it.
Parts of the microscope
When you place a transparent piece of onion skin under the microscope, you discover a grid of tiny compartments that are invisible to the naked eye. To see them clearly you need to hold the slide, light it and focus it. Which parts work together to make this magnification possible?
In the light microscope diagram, there are three parts labeled with "?". Write the name of each one and explain its function.
In the light microscope diagram, there are three parts labeled with "?". Write the name of each one and explain its function.
Name the two parts of the microscope that contain lenses and explain where you look through and where the specimen is placed.
Name the two parts of the microscope that contain lenses and explain where you look through and where the specimen is placed.
Explain what the microscope slide and the coverslip are and what each one is used for when preparing a specimen.
Explain what the microscope slide and the coverslip are and what each one is used for when preparing a specimen.
Carrying and putting away the microscope
A microscope is expensive and delicate. Carry it with two hands: one hand holds the arm and the other supports the base. Keep it upright and close to your body, and never swing it or drag the cord.
Describe how a microscope should be carried from the cabinet to the lab table without damaging it: which parts you hold and with how many hands.
Describe how a microscope should be carried from the cabinet to the lab table without damaging it: which parts you hold and with how many hands.
What must be done when finishing a microscope lab? Put these tasks in order and explain them: switching off the light, cleaning the lenses with the special lens paper,…
What must be done when finishing a microscope lab? Put these tasks in order and explain them: switching off the light, cleaning the lenses with the special lens paper, removing and disposing of the specimen, putting the microscope away, and washing your hands.
Total magnification
A microscope has an eyepiece lens of $10\times$ and objective lenses of $4\times$, $10\times$, and $40\times$. Calculate the total magnification achieved with each…
A microscope has an eyepiece lens of $10\times$ and objective lenses of $4\times$, $10\times$, and $40\times$. Calculate the total magnification achieved with each objective lens, and explain which one you should always start observing with and why.
A microscope has an eyepiece lens of $10\times$ and an objective lens of $20\times$. Calculate the total magnification. If you switch to a $40\times$ objective lens,…
A microscope has an eyepiece lens of $10\times$ and an objective lens of $20\times$. Calculate the total magnification. If you switch to a $40\times$ objective lens, explain what happens to the size of the image, to the area of the specimen you are able to see, and to the amount of light.
Field of view, light, and inverted images
When you switch to a higher-power objective lens, three things change. The image gets bigger, the field of view (the circle you see) gets smaller, and less light reaches your eye, so the image gets darker. If the image is too dark, open the diaphragm or turn up the light. If you can see only part of the specimen, center it on the stage. Always focus from lower to higher magnification.
The microscope image is inverted: upside down and reversed left to right. So the letter "b" looks like "q". If you move the slide to the right, the image seems to move to the left.
The microscope image is inverted. If you gently push the specimen to the right while looking, reason in which direction the image will seem to move in the eyepiece and…
The microscope image is inverted. If you gently push the specimen to the right while looking, reason in which direction the image will seem to move in the eyepiece and what practical consequence this has when you are following a moving microorganism.
With the $10\times$ objective lens (total magnification $100\times$), the field of view is about $1.5\ \text{mm}$ in diameter. If an onion cell is about…
With the $10\times$ objective lens (total magnification $100\times$), the field of view is about $1.5\ \text{mm}$ in diameter. If an onion cell is about $0.25\ \text{mm}$ long, estimate how many cells in a row would fit across the field of view, and explain what will happen to the diameter of the field of view when switching to the $40\times$ objective lens.
A classmate observes a slide under a microscope at $400\times$. The field of view with that objective lens measures $0.4\ \text{mm}$ in diameter, and the cell they are…
A classmate observes a slide under a microscope at $400\times$. The field of view with that objective lens measures $0.4\ \text{mm}$ in diameter, and the cell they are observing occupies approximately half of that diameter. Your classmate claims: "Since the microscope magnifies $400$ times and the cell occupies $0.2\ \text{mm}$, its actual size must be $0.2 \times 400 = 80\ \text{mm}$, it's almost eight centimeters long!".\na) Identify the conceptual error in their reasoning and explain the difference between actual size and magnified apparent size.\nb) Calculate the correct actual size of the cell in millimeters and micrometers ($1\ \text{mm} = 1\,000\ \mu\text{m}$).\nc) If you placed identical cells one after another in a row, how many would fit along one centimeter of a ruler?
Preparing a specimen
Put the steps for preparing an onion cell sample in order: (a) cover with the coverslip, taking care to avoid air bubbles, (b) place a thin layer of onion on the…
Put the steps for preparing an onion cell sample in order: (a) cover with the coverslip, taking care to avoid air bubbles, (b) place a thin layer of onion on the microscope slide, (c) add a drop of iodine solution (Lugol's solution) to stain, (d) remove excess liquid with filter paper.
Why are many samples stained before observing them under a microscope? Give an example of a stain used in the school lab.
Why are many samples stained before observing them under a microscope? Give an example of a stain used in the school lab.
To observe something under the light microscope it must be cut into very thin sections. Explain why the specimen must let light through and what happens if it is too…
To observe something under the light microscope it must be cut into very thin sections. Explain why the specimen must let light through and what happens if it is too thick.
Describe how you would prepare a wet mount of a hair or of a fiber from your sweater to view it under the microscope: what materials you would use and in what order you…
Describe how you would prepare a wet mount of a hair or of a fiber from your sweater to view it under the microscope: what materials you would use and in what order you would act. What is the difference between that homemade preparation and a permanent laboratory slide?
Focusing correctly
Why must you focus first using the lowest power objective lens and reserve the fine focus knob for higher power objective lenses? Explain what might happen if you use…
Why must you focus first using the lowest power objective lens and reserve the fine focus knob for higher power objective lenses? Explain what might happen if you use the coarse focus knob with the $40\times$ objective lens.
You have focused a cell with the $10\times$ objective lens and switch to the $40\times$ one: the image is left slightly blurry and darker. Explain which focus knob you…
You have focused a cell with the $10\times$ objective lens and switch to the $40\times$ one: the image is left slightly blurry and darker. Explain which focus knob you must use to refocus (and which one you must not touch) and which lighting adjustment can improve the image.
When observing a sample, the image looks very dark and only a fragment of the specimen can be seen. State two possible causes for each problem and what you would do to…
When observing a sample, the image looks very dark and only a fragment of the specimen can be seen. State two possible causes for each problem and what you would do to correct them without changing the objective lens.
Lab safety and good practice
Before any lab, think about safety. Stains and biological samples need care, and your lab notebook is where you record what you see.
Why? Stains such as iodine can irritate your eyes and stain skin and clothes, and some samples may carry germs. Broken coverslips are very sharp.
Name three safety rules for working with biological samples and stains in the school lab and explain the reason for each one.
Name three safety rules for working with biological samples and stains in the school lab and explain the reason for each one.
During a lab activity observing cheek cell specimens stained with methylene blue, a group can only see shapeless blue smudges. Suggest at least three errors they might…
During a lab activity observing cheek cell specimens stained with methylene blue, a group can only see shapeless blue smudges. Suggest at least three errors they might have made when preparing the sample and explain how you would repeat the experiment to get good results.
You want to check whether the cells in the growth zone of an onion root are smaller than those in the bulb. Design the lab activity: which two specimens you will…
You want to check whether the cells in the growth zone of an onion root are smaller than those in the bulb. Design the lab activity: which two specimens you will prepare, how you will estimate the size of the cells using the field of view, which data table you will use, and how many cells you will measure so that the comparison is reliable.
When observing the letter "e" cut out from a newspaper, the microscope shows it rotated 180° (upside down and backward). Predict how the letter "p" will look before…
When observing the letter "e" cut out from a newspaper, the microscope shows it rotated 180° (upside down and backward). Predict how the letter "p" will look before viewing it, explain why this inversion happens, and describe how you would test your prediction.
Unit challenge. Imagine that you are collaborating with your state's science museum by designing an informative sheet on microscopic life in the home. 1. Search for real…
Unit challenge. Imagine that you are collaborating with your state's science museum by designing an informative sheet on microscopic life in the home.
-
Search for real samples: Access the free image bank of the NIH (National Institutes of Health) or the Smithsonian photo library. Download two real micrographs: a bacterium (Lactobacillus from yogurt) and a plant cell (onion epidermis). Record the magnifications and the indicated scale bar.
-
Measurement and analysis: Measure the cell in the photo with your ruler and, applying the graphic scale, calculate its actual size in micrometers ($1\text{ }\mu\text{m} = 0.001\text{ mm}$). Prepare a comparative table that includes: calculated actual dimensions (e.g. $1.8\text{ }\mu\text{m}$ compared to $120.5\text{ }\mu\text{m}$), cell type (prokaryotic or eukaryotic), and the presence or absence of a visible nucleus, cell wall, and cytoplasm.
-
Final report: Write a reasoned proposal (maximum $1$ page) explaining why the cell is the unit of life in both organisms, why they differ under the microscope, and what advantages this instrument offers for studying them.
Assessment criteria:
-
Rigorous identification and classification of prokaryotic and eukaryotic cellular structures.
-
Accuracy in calculating actual cell size from the microscope scale.
-
Conceptual clarity when justifying that the cell is the functional and structural unit of living things.