Microscopes and separating cell components Flashcards
What is the magnification
The degree to which an object is magnified (How many times bigger the image is)
What is the resolution
and What microscope has the higher resolution and why
resolution = Ability to distinguish between two points (Dependent ton the wavelength of the radiation used to view to object)
Electron microscopes electron beam have a smaller wavelength so it has a higher resolution than the light microscopes light beam
properties of a transmission electron microscope
- Specimens cant be alive must be viewed in a vacuum
- specimens must be thin and stained
- parts of the specimen absorbs the beam of electrons appearing darker than parts that don’t.
- Resolving power 0.1nm
- Artifacts may appear - part of the micrograph but not part of the natural object. Results from preparation process
Properties of a scanning electron microscope
- electrons don’t penetrate the specimen, so the sample doesn’t need to be thin
- The electrons are scattered by the sample, allowing a 3d image to be formed
- Resolving power is 20nm
what is the first step on separating cell components
- Broken cells/tissues are placed in a cold, buffered, isotonic solution
- Then put in a homogeniser
why are the cells/tissue put in cold, buffered, isotonic solution
cold = stopping enzyme reactions
buffered = keeps PH neutral and means the enzymes don’t denature and keeps the membrane
isotonic = same water concentration meaning no osmosis
what happens in the 2nd stage of separating cell components
the homogenate is filtered to remove unbroken cells and then be placed in a centrifuge on a low speed
what happens in the 3rd stage of seperating cell components
Once the centrifuge has stopped the denser organisms will go to the bottom of the tubes becoming pellets
- the supernatant is removed and re-spun at a higher speed
repeat at higher speeds
What is the order of density and how many times the supernatant will have to be centrifuged
- nuclei
- mitochondria and chloroplast
- lysosomes
- endoplasmic reticulum
Why do the specimens need to be in a vacuum, thin and stained for a transmission microscope
Vacuum- electrons are absorbed by air
Thin- so the electrons can pass through
Stained - because the image is black and white
How do you convert millimetres into micrometres and vise versa
mm into um = x1000
um into mm = /1000