Chapter 3 Flashcards
What is a buffered solution used for?
To resist small changes in pH which denature proteins in the membranes of cell organelles
Why should a buffered solution for cell fractionation be cold?
To reduce the effect of enzymes that would digest the cell organelles
What does isotonic mean for a cell?
Means that the solution outside the cell is the same concentration as the in solution inside the cell. Means that they have the same water potential
What does hypertonic mean for a cell?
Means that the solution outside the cell is a higher concentration than the in solution inside the cell. Means that the cell has a low water potential.
What does hypotonic mean for a cell?
Means that the solution outside the cell a lower concentration than the in solution inside the cell. Means that the cell has a high water potential
Define homogenate
A fluid in which everything is evenly distributed
What is the effect of ultracentrifugation?
During spinning the heaviest organelles will separate out and the the lightest organelles will separate out last
State the equation for magnification
Magnification = size of image / size of object
Define resolution
The minimum distance apart that 2 objects can be in order for them to appear as 2 distinct items
The greater the resolution the greater the ________ of the image
Clarity
What controls the resolving power of a microscope?
The wavelength size and the form of radiation used
What is cell fractionation?
The process where cells are broken up and the different organelles they contain are separated out
What is ultracentrifugation?
The process in which the fragments in a filtered homogenate are separated out in a machine called a centrifuge
Describe the process of ultracentrifugation in animal cells
The homogenate is spun at a low speed and the heaviest organelles are forced to the bottom via centrifugal force. The supernatant is then removed and placed into a different tube and spun faster than before to separate out mitochondria which are then forced to the bottom This process is continued for more organelles
Why do light microscopes have poor resolutions?
Due to the long wavelength of light
When was the electron microscope developed?
In the 1930s
Name 2 advantages of electron microscopes over light microscopes
Higher resolving power due to a short electron wavelength (0.1nm) Electrons can be focused using electomagnets
What does TEM stand for?
Transmission electron microscope
What is the resolving power of a tem?
0.1nm
Why might the resolution of a tem be altered?
A high electron beam could destroy the specimen The specimen may be prepared wrong
What kind of image is produced by a tem? What is this projected on?
2D A fluorescent screen
Does a TEM make a colour image?
No
How does a TEM work?
An electron beam is passed through a v thin layer of specimen and some parts absorb electrons and some pass them through making darker and lighter spots which make the image
List 5 limitations of a TEM
Vaccuum needed - specimen is not alive Complex staining process Image not coloured Specimen must be extremely thin - not whole view Artefacts may appear and confuse or distort the image
How does an SEM work?
A beam of electrons is sent to the specimen but instead of penetrating the object the electrons are scattered and the pattern of the scatter and the secondary electrons make the image
What is a limitation of a SEM that a TEM doesnt have
The resolving power is lower - 20nm not 0.1nm
What is a limitation that a TEM has that an SEM doesnt?
The specimen has to be thin and the image is 2D
What is the the magnification of a Light microscope TEM SEM
400x 150,000x 100,000x
When is a vaccum required ?
SEM TEM
What is the resolving power of a light microscope?
200nm
Why do light microscopes have poor resolutions?
Due to the long wavelength of light
When was the electron microscope developed?
In the 1930s
Name 2 advantages of electron microscopes over light microscopes
Higher resolving power due to a short electron wavelength (0.1nm) Electrons can be focused using electomagnets
What does TEM stand for?
Transmission electron microscope
What is the resolving power of a tem?
0.1nm
Why might the resolution of a tem be altered?
A high electron beam could destroy the specimen The specimen may be prepared wrong
What kind of image is produced by a tem? What is this projected on?
2D A fluorescent screen
Does a TEM make a colour image?
No
How does a TEM work?
An electron beam is passed through a v thin layer of specimen and some parts absorb electrons and some pass them through making darker and lighter spots which make the image
List 5 limitations of a TEM
Vaccuum needed - specimen is not alive Complex staining process Image not coloured Specimen must be extremely thin - not whole view Artefacts may appear and confuse or distort the image
How does an SEM work?
A beam of electrons is sent to the specimen but instead of penetrating the object the electrons are scattered and the pattern of the scatter and the secondary electrons make the image
What is a limitation of a SEM that a TEM doesnt have
The resolving power is lower - 20nm not 0.1nm