Methods Of Studying Cells Flashcards
What is magnification
It’s how much bigger the image is than the specimen
What is resolution
Is how detailed the image. It’s how well a microscope distinguished between 2 points that are close together
What is the formula for magnification
Magnification = size of image / size of real object
M=I/a
Before cell fractionation can take place, what must the solution be like
Cold- to reduce enzyme activity that might break down the organelles
Is of the same water potential as the tissue- to prevent organelles bursting or shrinking as a result of osmotic gain or loss of water
Buffered - so that the pH does not fluctuate. A change in pH could alter the structure of the organelles or affect the functioning of enzymes
What’s the first stage in cell fractionation
Homogenisation - cells are broken up by a homogeniser, this releases the organelles from the cell. It is then filtered to remove any complete cells and large pieces of debris
What is the second stage of cell fractionation
Ultracentrifugation- is the process by which the fragments in the filtered homogenate are separated in a machine called a centrifuge.
What’s the process of ultracentrifugation
The tube of filtrate is placed in the centrifuge and spun at slow speed.
The heaviest organelles, thin nuclei, are forced to the bottom of the tube, where they form a pellet
The fluid at the top of the tube (supernatant) is removed leaving just the sediment of nuclei
The supernatant is transferred to another tube and spun in the centrifuge at a faster speed than before
The next heaviest organelles, the mitochondria, are forced to the bottom of the tube
The process is continued in this way so that, at each increase in speed, the next heaviest organelle is sedimented and separated out