cell fractionation

what must be done in order to look at specific organelles?

they must be separated, which is done by cell fractionation

First step

homeogenation

homeogeniasation

grind up in a blender to break cell surface and release organelles

3 conditions and why?

-cold solution- reduce enzyme activity

-isotonic-water potential remains same, reduce osmosis effect

-buffer- constant ph to prevent denaturing of proteins

second step

filter to remove debris through a gauge

third step

ultracentrifuguration

ultracentifiguration

the cell fragments are transferred to a test tube to form a sediment pellet

spin at a low speed

the densest organelles sink at the bottom, sedemented pellet, the rest of the organelles are left in a solution (supernant)

the supernant is drained and poured into another tube and spun at a higher speed

repeat until every organelle is separated based on their densities

differential centrifiguration

spins at a faster speed

centrifuge

Device that separates organelles based on their density

densest to less dense

nuclei, mitochondria, er, ribosomes

what force does the centrifuge create?

centrifugal force

what did ultracentrifiguration develop?

advances in biological knowledges. Detailed structure and function of organelles