cell fractionation and ultracentrifugation

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Why is cell fractionation used?

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used to isolate different organelles so that they can be studied

- enables individual organelle structures and functions to be studied

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explain why fractionated cells are kept in a cold, buffered, isotonic solution

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cold: to reduce the activity of enzymes that break down organelles

buffered: solution has to have a pH buffer to prevent damage to the organelles

isotonic: organelles must be the same water potential as the solution to prevent osmosis

- osmosis could cause the organelles to shrivel or burst

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4 Terms

1
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Why is cell fractionation used?

used to isolate different organelles so that they can be studied

- enables individual organelle structures and functions to be studied

2
New cards

explain why fractionated cells are kept in a cold, buffered, isotonic solution

cold: to reduce the activity of enzymes that break down organelles

buffered: solution has to have a pH buffer to prevent damage to the organelles

isotonic: organelles must be the same water potential as the solution to prevent osmosis

- osmosis could cause the organelles to shrivel or burst

3
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describe what happens during cell fractionation

homogenisation:

1) the cells must be broken open in the cold, buffered, isotonic solution

(can be done using a blender or by crushing them)

filtration

2) solution is filtered through a gauze to remove large cell debris or tissue debris

ultracentrifugation

3) the filtered solution is spun at high speed in a centrifuge

- this separates organelles according to their density

4) this process is repeated several times

- each time you remove the supernatant (the liquid containing all the organelles that have to be separated) and leave behind the pellet (the organelle that has been isolated)

4
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State the order of sedimentation of organelles during differential centrifugation.

most dense to least dense:

1. nucleus

2. mitochondria

3. lysosomes

4. RER

5. plasma membrane

6. SER

7. ribosomes