cell fractionation and ultracentrifugation

purpouse

to separate different organelles to study structure and function

step 1

tissue is placed in cold, buffered, isotonic solution

cold solution

reduce enzyme activity, preventing organelle digestion

isotonic solution

prevents bursting or shrinking due to osmosis

buffered solution

so the ph doesn’t fluctuate and prevents denaturing

homogenation

step 2

use a blender to break open cells to release organelles

filter to remove debris

ultracentrefugation

step 3

spin on low speed

organelles separate by mass

remove supernatant and spin faster

repeat