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Cell Fractionation Flashcards

(9 cards)

1
Q

Define cell fractionation.

A

The process where cells are broken up (lyses) and the different organelles within the cell are separated out.

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2
Q

What is the tissue placed in before cell fractionation can begin?

A
  • A cold, buffered solution with the same water potential as the tissue.
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3
Q

Why is the isolation medium in cell fractionation a **cold*, buffered solution with the same water potential as the tissue?

A
  • Cold to keep Enzyme activity low and prevent hydrolytic enzymes damaging organelles.
  • Buffered to keep the pH stable to all the organelles and their enzymes to function properly.
  • The same water potential to make upsurge that water does not energ or leave the organelles by osmosis, so they don’t swell or shrink and get damaged.
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4
Q

What are the three steps of cell fractionation called?

A
  • Homogenization
  • Filtration
  • Ultracentrifugation
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5
Q

Describe the homegenization stage of cell fractionation.

A
  • This breaks open the cells, usually done by either vibrating the cells or grinding them up in a homogeniser (blender).
  • This releases organelles from the cells.
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6
Q

Describe the filtration stage of cell fractionation.

A
  • The resulting fluid (homogenate) is filtered to remove any debris (whole cells or large bits of remaining tissue).
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7
Q

Describe the ultracentrifugation stage of cell fractionation.

A

The fragments in the filtered homogenate are separated in a centrifuge at increasing speeds.

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8
Q

What is the supernatant?

A
  • Smaller and less dense components.
  • Used if performing repeated centrifugation.
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9
Q

What is the pellet?

A
  • Larger and more dense components.
  • Removed before repeated centrifugal ion.
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