Methodology Flashcards
light microscopy
thin sections of tissue; can e used on live or fixed and stained cells; white light or transmitted light
resolution
amount of detail/info retained from the specimen upon magnification
fluorescent light microscopy
same limit of resolution as white light microscopy
can observe fluorophores even in live cells
high contrast
FRET
Fluorescent Resonance Energy Transfer
good for determining:
1. how close in a cell two molecules are
2. to follow the activation of an enzyme in situ
laser scanning confocal microscopy
similar to fluorescent microscopy, but uses lasers as illumination and can only visualize one focal plane at a time
larger specimens
FRAP
Fluorescence recovery after photobleaching
scanning electron microscopy (SEM)
reveals specimen topography
specimens are dehydrated then metal coated
max resolution of SEM
5nm
transmission electrom microscopy (TEM)
very high resolution images
problem with TEM
potential for artifacts
resolution of TEM
0.1nm-2nm
cell culture
growing cells outside of the organism
differential centrifugation
homogenize cells and isolate some organelles which can then be separated from one another through sub-cellular fractionation