Examining Cells Under the Microscope
Viewing cells under the microscope
- Many biological structures are too small to be seen by the naked eye
- Optical microscopes are an invaluable tool for scientists as they allow for tissues, cells and larger organelles to be seen and studied
- Light is directed through a thin layer of biological material that is supported on a glass slide
- This light is focused through several lenses so that an image is visible through the eyepiece
- Getting a visible image requires a very thin sample of biological tissue because light has to pass through the sample and into the lenses of the microscope
- The most common specimens to observe under a light microscope are cheek cells (animal cells) and onion cells (plant cells)
- A stain is often used to ensure cell structures are clearly visible under the microscope
General method
- Specimens must be prepared on a microscope slide to be observed under a light microscope
- This must be done carefully to avoid damaging the biological specimen and the structures within it
- Preparing a slide using a liquid specimen:
- Add a few drops of the sample to the slide using a pipette
- Cover the liquid/smear with a coverslip and gently press down to remove air bubbles
- Wear gloves to ensure there is no cross-contamination of foreign cells
- Preparing a slide using a solid specimen:
- Use scissors to cut a small sample of the tissue
- Peel away or cut a very thin layer of cells from the tissue sample to be placed on the slide (using a scalpel or forceps)
- Some tissue samples need to be treated with chemicals to kill/make the tissue rigid
- Gently place a coverslip on top and press down to remove any air bubbles
- A stain may be required to make the structures visible depending on the type of tissue being examined
- Commonly used stains include methylene blue to stain cheek cells and iodine to stain onion cells
- Take care when using sharp objects and wear gloves to prevent the stain from dying your skin
- When using an optical microscope always start with the lowest power objective lens:
- It is easier to find what you are looking for in the field of view
- This helps to prevent damage to the lens or coverslip in case the stage has been raised too high
- Preventing the dehydration of tissue:
- The thin layers of material placed on slides can dry up rapidly
- Adding a drop of water to the specimen (beneath the coverslip) can prevent the cells from being damaged by dehydration
- Unclear or blurry images:
- Switch to the lower power objective lens and try using the coarse focus to get a clearer image
- Consider whether the specimen sample is thin enough for light to pass through to see the structures clearly
- There could be cross-contamination with foreign cells or bodies
Using a microscope diagram
Light microscopes have a lens in the eyepiece which is fixed and two or three objective lenses of different powers
Viewing plant tissue
- An ideal tissue is the onion epidermis (found between the layers of onions) because it forms a layer just one cell thick
- Being a non-photosynthetic tissue, onion epidermis is not green as it does not contain any chloroplasts
Apparatus
- The key components of an optical microscope you will need to use are:
- The eyepiece lens
- The objective lenses
- The stage
- The light source
- The coarse and fine focus
- Other apparatus used:
- Forceps
- Scissors
- Scalpel
- Coverslip
- Slides
- Pipette
- Iodine solution
Viewing onion cells under the microscope diagram
Care must be taken to avoid smudging the glass slide or trapping air bubbles under the coverslip
Viewing animal tissue
- Human cheek cells are a good choice for examination under the light microscope because they are:
- Plentiful
- Easy to obtain safely
- Can be obtained without an overly intrusive process
- Relatively undifferentiated and so will display the main cell structures
Safety considerations
- Do not perform the sampling on a person who has a cold, cough, throat infection etc.
- To avoid spreading the infection to others
- Concentrated methylene blue is toxic if ingested
- Wear gloves and do NOT allow children to handle methylene blue solution or have access to the bottle of solution
- Wear gloves and do NOT allow children to handle methylene blue solution or have access to the bottle of solution
Apparatus
- Glass microscope slides
- Cover slips
- Paper towels or tissue
- Staining solution
- Methylene blue solution
- 0.5% to 1%
- Dilute according to concentration of the stock solution
- 0.5% to 1%
- Methylene blue solution
- Plastic pipette or dropper
- Sterile, individually packed cotton wool buds or swabs
Method
- Brush teeth thoroughly with normal toothbrush and toothpaste
- This removes bacteria from teeth so they don't obscure the view of the cheek cell
- Take a clean, sterile cotton swab and gently scrape the inside cheek surface of the mouth for 5-10 seconds
- Smear the cotton swab on the centre of the microscope slide for 2 to 3 seconds
- Add a drop of methylene blue solution
- Place a coverslip on top
- Lay the coverslip down at one edge and then tilt it down flat
- This reduces bubble formation under the coverslip
- Lay the coverslip down at one edge and then tilt it down flat
- Absorb any excess solution by allowing a paper towel to touch one side of the coverslip.
- Place the slide on the microscope, with 4 x or 10 objective in position and find a cell
- Then view at higher magnification to reveal more detail
- Methylene blue stains negatively charged molecules in the cell, including DNA and RNA
- This causes the nucleus and mitochondria appear darker than their surroundings
- The cells seen are squamous epithelial cells from the outer epithelial layer of the mouth
Viewing cheek cells under the microscope diagram
Parts of the cell that can be seen with a light microscope
- Nucleus
- Mitochondria
- Cell membrane
- Cytoplasm
Parts of the cell that cannot be seen with a light microscope
- Ribosomes*
- Endoplasmic reticulum*
- Golgi*
- Details of the nucleus, mitochondria and cell membrane
* these are parts of the cell that you don't need to know the names of, but they are included here as examples of very small structures within the cell
Examiner Tip
Remember that a cell is always a 3-dimensional object. If you search around your microscope's field of view you may find a cell squashed up against the coverslip and will be able to see its 3-D structure in the background.