Analysis of Cell Components

Magnification - how much bigger the image is than the specimen (the sample your looking at)

Formula for magnification = length of image/length of specimen

ie. if you magnified image that's 5mm wide and your specimen is 0.05mm wide the magnification is: 5/0.05= times 100

Resolution - how detailed the image is. More specifically, it's how well a microscope distinguishes between two points that are close together. If a microscope lens can't seperate two objects, then increasing the magnificationwon't help.

Light Microscopes;

• They use light.
• They have lower resolution than electron microscopes.
• They have a maximum resolution of about 0.2 micrometres (um).
• The maximum useful magnification of a light microscope is about times 1500.

Electron Microscopes;

• They use electrons instead of light to form an image.
• They have a higher resolution than light microscopes so give a more  detailed image.
• They have a maximum resolution of about 0.0001 micrometres (about 2000 times higher than light microscopes.)
• The maximum useful magnification of an electron microscope is about 1,500,00.

Transmitting Electron Microscopes (TEMs);

• TEMs use elctromagnets to focus a beam of electrons, which is then transmitted through the specimen.
• Denser partsof the specimen absorb more electrons, which make them look darker on the image you end up with.
• TEMs are good because they give high resolution images.
• But they can only be used on thin specimens.

Scanning Electron Microscopes (SEMs);

• SEMs scan a beam of electrons across the specimen.
• This knocks of electrons from the specimen, which are gathered in a cathode ray tube to form an image.
• The images you end up with show the surface of the specimen and they can be 3D.
• But they give lower resolution images than TEMs.

1) Homogenisation -  breaking up the cells. e.g. by vibrating the cells by grinding the cellsup in a blender. Thsi breaks up the plasma membrane and releases the organelle into the solution.

2) Filtration- gettng rid of the big bits. The solution is filtered though a gauze to seperate any large cell debris or tissue debris. The organelles are much smaller than the debris, so they pass through.

3) Ultracentrifugation - Seperating the organelles.

- The cell fragments are poured into a tube. This tube is put into a cetrifuge and spun at a low speed. the heaviest organelles, like nuclei, get flung to the bottom - the pellet. The rest of the organelles stay suspended in the fluid above the sediment - the supernatant.

- The supernatant is drained off, poured into another tube, and spun in a centrifuge at a higher speed.