Pack 2 - Cell Structure Flashcards
Define resolution.
The minimum distance apart two objects can be in order for them to be distinguishable as two separate objects.
What determines the resolution of a microscope?
The wavelength of radiation being used. (e.g the wavelength of light)
What is the approximate resolution of a light microscope?
0.2μm
Why is the resolving power of an electron microscope higher than a light microscope?
The wave length of electrons is shorter than the wavelength of light.
What is cell fractionation?
The process where cells are broken up and the organelles they contain are released and separated out.
What THREE conditions must a tissue be under before cell fractionation can take place and WHY?
- Cold - To reduce enzyme activity that may break down ORGANELLES.
- Isotonic (same Ψ as tissue) - to prevent bursting or shrinking of ORGANELLES due to osmosis.
- Buffered (same pH as tissue) - to prevent enzymes denaturing or the changing in the structure ORGANELLES.
Describe cell fractionation in 4 steps.
- Homogenisation - cells are broken up by a blender into a homogenate releasing organelles.
- Filtration - whole cells/debris removed
- Ultracentrifugation - homogenate is spun in a CENTRIFUGE at a low therefore the densest organelles fall to the bottom.
- Repeat - the supernatant is poured of and the process is repeated on the supernatant at higher speeds to obtain less dense organelles.
What is the fluid at the top of a centrifuge tube called after centrifugation?
Supernatant
Order the following organelles by increasing speed of centrifugation.
• Mitochondria
• Nuclei
• Lysosomes
- Nuclei
- Mitochondria
- Lysosomes
How are electrons focused in an electron microscopes?
Using electromagnets (because electrons are negatively charged).
What is an important condition needed for all electron microscopes and why?
A near vacuum - because electrons are deflected and absorbed by air molecules.
What kind of image does a transmission electron microscope (TEM) produce?
2D, Black and White
What is the approximate resolution of a transmission electron microscope (TEM)?
0.1nm
Why can the maximum resolving power of a transmission electron microscope (TEM) no always be produced?
- Difficulties preparing the specimen.
* high energy electron beams required would destroy the specimen.
What are 4 main limitations of a TEM?
- The whole system must be in a near vacuum therefore living specimens cannot be observed.
- A complex staining process is required.
- The specimen must be extremely thin to allow electrons to pass through.
- The image may contain artefacts due to the way the specimen is prepared.
What kind of image does a scanning electron microscope (SEM) produce and how?
3D - Black and white
Rather than penetrating the sample, a SEM passes a beam of electrons over the sample in a regular pattern. The electrons are scattered depending on the surface. A computer builds a 3D image based on the scattering of electrons.
What are the advantages of a SEM over a TEM?
- Specimen does not have to be thin.
* 3D image
What is the approximate resolution of a scanning electron microscope (SEM)?
20nm
Name the 5 components of the nucleus and their roles.
- Nuclear envelope - controls the entry and exit of materials; contains reactions taking place in the nucleus.
- Nuclear pores - allows passage of large molecules in and out of the cell e.g. mRNA
- Nucleoplasm
- Chromosomes - consist of protein bound linear DNA
- Nucleolus - manufactures ribosomal RNA and assembles ribosomes.