9. Cell Fractionation Flashcards
What is Cell Fractionation?
CELL FRACTIONATION:
PROCESS USED TO SEPARATE CELLULAR COMPONENTS WHILST PRESERVING INDIVIDUAL FUNC-TIONS OF EACH COMPONENT TO STUDY THE ORGANELLES PRESENT INSIDE CELLS.
ORGANELLES SEPERATED BY PHYSICAL PROCESSES – BREAKING UP CELLS - SEPARATE INDVIDUAL ORGANELLES ACCORDING TO SIZE
Method/ Process of Cell fractionation?
Method/process:
- Obtain sample of tissue w/ cells for study
- Tissue cut to small pieces
-Homogenisation : in blender to break up cells walls/membranes - free contents:
o An isotonic and pH buffer solution are ground, minced and chopped with the sample tissue to stop osmotic damage
-Filtration
o To removes debris
Purification : filtrate is spun in very powerful centrifuge
o By Centrifugation
o The Nuclei (largest organelle) the first fraction (usually)
o After first faction mitochondria/ chloroplasts
o After mitochondria/ chloroplasts RER req higher spinning /tiny ribosomes require the most spinning to get them to sediment (300,000g)
What is Centrifugation?
o CENTRIFUGATION:
PROCESS SEPARATING SOLIDS FROM LIQUIDS/ LIQUIDS OF DIF-FERENT DENSITIES FROM EACH OTHER BY USING CENTRIFUGAL FORCE. PROCESS OCCURS INSIDE A CENTRIFUGE.
