Histo Methods
Terms
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copy deck
- resolving power of eye vs. light microscope vs. electron microscop
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250 microns
0.25 microns
0.001 microns - wavelenght of light vs. electrons
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0.5-0.55mircons
0.004 nm - Azan stains
- stains collagen blue
- iron hematoxylin stains
- stains mitchondira
- silver stains
- stains Golgi and reticular fibers (collagen type III)
- toluidine blue stains
- stains mast cells and RNA
- Verhoeff stains
- stains protein black
- what does a PAS (periodic acid shift) do?
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histochemistry. periodic acid oxidizes glycols to adehydes. Schiffs reagent then stains the alehydes magenta. so used to see carbs (glycols=adjacent carbons bearing -OH (hydroxyl) groups)
is a specific molecule present? - Enzyme histochemistry
- used to pinpoint location of specific enzymes in cell. (eg ATPase). makes insoluble, colored/visible product that preciptates at site where enzyme located. if not visible, can trap and precipitate with Chromagen (colored)
- Immunoflourescence
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used to pinpoint location of specific molecules in cell
absorubs shot wavelength, emits longer one
seconadry antibody (Indirect) often used
DAPI-->DNA blue (direct)
FITC+ antibody -->MTs (indirect) - Autoradiography
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used to pinpoint location of specific gene being expressed
eg. 1. radiolabel AAor nucleotides
2. In Situ hybridization: used to detect location of specific mRNA, radiolabel cDNA/anti-sense RNA which hybridize with mRNA
3. ligand-bindind assays