Principles of optical microscopes
Simple convex glass lenses used in pairs in a compound light microscope to focus an object at a short distance by 1st lens, then magnified by 2nd lens
Pros and cons of optical microscope
Pros:
- Cheap
- Images in colour
- No training required
- Live specimens
Cons:
- Low magnification
- Low resolution
- 2D images
Principles of transmission electron microscope
Pros and cons of transmission electron microscope
Pros:
- High resolution images
- High magnification
- Visible internal structures
Cons:
- Expensive
- Training is required
- No colour images
- 2D images
- Only thin specimens
Principles of scanning electron microscope
Pros and cons of scanning electron microscope
Pros:
- 3D images
- High magnification
- High resolution
- Thick specimens
Cons:
- Expensive
- Training is required
- No colour images
How do you prepare a slide for an optical microscope
What is the difference between magnification and resolution?
Magnification: Increasing the size of an image. Up until the limit of resolution, an increase in magnification = an increase in detail
Resolution: minimum mistaken apart that two objects can be for them to appear separate items
What is the formula to calculate magnification
Can you describe the principles of cell fractionation and ultracentrifugation in separating cell components?
Why is a cold, isotonic buffer needed
Cold - to reduce enzyme activity that could break down organelles
Isotonic - same water potential as tissue sample - to prevent water moving in or out of the cells by osmosis, causing lysis
Buffered - to prevent changes in pH which could affect/denature enzymes
How are organelles separated out during centrifugation?
They are separated in order of mass and the order is usually
Nuclei
Mitochondria
Lysosomes
Endoplasmic reticulum
Ribosomes
Distinguishing features of eukaryotic cells
general structure of animal cell
General plant structure
Same as animal
- Chloroplast
- Cell vacuole
- Cell wall
Cell surface membrane
What does the nucleus consist of
Nuclear envelope:
- Double membrane
- Has nuclear pores which allow substances e.g. mRNA to move between nucleus and cytoplasm
Nucleolus:
- Makes ribosomes
Nucleoplasm: granular jelly like material that makes up the bulk of the nucleus
Protein bound, linear DNA:
- Chromatin = condensed
- Chromosome = highly condensed
Ribosomes
Two subunits = large subunit and small subunit, each of which contains ribosomal RNA and protein
- Not surrounded by a membrane but can be attached to RER
- Site of protein synthesis - translation
smooth endoplasmic reticulum
Rough endoplasmic reticulum
Golgi apparatus and vesicles
Golgi apparatus = flattened membrane sacs
- Modifies/processes protein from RER
- e.g. protein + carbohydrate –> gylcoproteins
- Packages them into Golgi vesicles
- Produces lysosomes (a type of Golgi vesicle)
Golgi vesicle = small membrane sac
- Transports proteins / lipids to their required destination
- e.g. to the cell-surface membrane
Lysosomes
Mitochondria
Components of the mitochondria:
- Outer membrane
- Cristae - inner membrane fold
- Matrix, containing: small / 70s ribosomes, circular DNA
Chloroplasts
Components of the chloroplast:
- Double membrane
- Stroma, containing: Thylakoid membrane, Small 70s ribosomes, Circular DNA, Starch granules / lipid droplets
- Lamella - thylakoid linking grana
- Grana - stacks of thylakoid