Cell Fractionation

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  • Created by: Meg_
  • Created on: 02-10-13 15:20

Cell Fractionation

Cell Fractionation:

Process of breaking down tissues/cells and separating out organelles.

Firstly, the tissue is put in a COLD, ISOTONIC and BUFFERED solution.

Cold to reduce enzyme activity that could break down the organelles.

Isotonic to prevent water movement.

Buffered to maintain a constant pH.

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Homogenisation

Then the tissue is HOMOGENISED in a HOMOGENISER which breaks down the cell. This process is HOMOGENISATION.

Usually done in a blender.

 

 

 

Afterwards the HOMOGENATE is filtered to remove debris.

Small organelles pass through but cell debris doesn't.

We are then left with the FILTRATE.

 

 

 

 

 

 

 

 

 

 

 

 

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Ultragentrifugation

The filtrate is spun in an ULTRACENTRIFUGE.

This is ULTRACENTRIFUGATION.

Heavy organelles fall to the bottom (SEDIMENT).

Lighter organelles form the SUPERNATANT which is separated and spun again at a faster speed.

This process continues until you are left with the organelles you want.

Cells separated out by speed (heaviest first):

Nuclei

Mitochondria

Lysosomes

Endoplasmic Reticulum

Ribosomes

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