Cell Fractionation
The process by which cells are broken up
and the organelles are isolated to be
investigated.
Ultracentrifugation Homogenisation Preparation
Step
Reason
Make sure
the 2 last
boxes are
larger than
the others!
Step
Reason
Tissue placed in a…
1) Cold
1)Prevent enzyme activity
2) Isotonic
2)Prevent osmosis
3) Buffered
3) maintain correct and
constant pH
…solution
Step
Reason
Tissue placed in a…
•
Blender or pestle • Break cells up
•
• Remove whole cells
Fluid extracted
(homogenate) and and debris
filtered
Step
Homogenate placed
into an
ultracentrifuge
1. Spun at low speed
(1000 x g)
2. Sediment in base,
supernatant on top
3. Extract supernatant
4. Spun at medium
speed (4000 x g)
5. Extract supernatant
6. Spun at high speed
(16000 x g)
Reason
1. Create a density gradient
2. Collect nuclei in sediment
3. Collect remaining
organelles
4. Collect mitochondria in
sediment
5. Collect remaining
organelles
6. Collect E.R, lysosomes
and ribosomes.
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,