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Cell Fractionation

Cell Fractionation. The process by which cells are broken up and the organelles are isolated to be investigated. Step. Reason. Preparation. Homogenisation. Make sure the 2 last boxes are larger than the others!. Ultracentrifugation. Reason Prevent enzyme activity Prevent osmosis

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Cell Fractionation

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  1. Cell Fractionation The process by which cells are broken up and the organelles are isolated to be investigated.

  2. Step Reason Preparation Homogenisation Make sure the 2 last boxes are larger than the others! Ultracentrifugation

  3. Reason • Prevent enzyme activity • Prevent osmosis • maintain correct and constant pH Step Tissue placed in a… • Cold • Isotonic • Buffered …solution

  4. Reason • Break cells up • Remove whole cells and debris Step Tissue placed in a… • Blender or pestle • Fluid extracted (homogenate) and filtered

  5. Reason • Create a density gradient • Collect nuclei in sediment • Collect remaining organelles • Collect mitochondria in sediment • Collect remaining organelles • Collect E.R, lysosomes and ribosomes. Step Homogenate placed into an ultracentrifuge • Spun at low speed (1000 x g) • Sediment in base, supernatant on top • Extract supernatant • Spun at medium speed (4000 x g) • Extract supernatant • Spun at high speed (16000 x g)

  6. Bingo! Draw a 3 x 3 grid and write in 9 of the following words: Lysosome, endoplasmic reticulum, mitochondria, golgi apparatus, ribosomes, enzymes, nuclei, isotonic, buffered, cold, homogenate, low speed, high speed, ultracentrifugation, cell fractionation,

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