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Team:Newcastle/Scanning EM
From 2010.igem.org
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Swoodhouse (Talk | contribs) (→Fixation) |
Swoodhouse (Talk | contribs) (→Dehydration) |
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==Dehydration== | ==Dehydration== | ||
| - | 25% ethanol: 30 mins. | + | *25% ethanol: 30 mins. |
| - | 50% ethanol: 30 mins. | + | *50% ethanol: 30 mins. |
| - | 75% ethanol: 30 mins. | + | *75% ethanol: 30 mins. |
| - | 100% ethanol: 1 hr. | + | *100% ethanol: 1 hr. |
| - | 100% ethanol: 1 hr. | + | *100% ethanol: 1 hr. |
Once in 100% ethanol, samples are dried using a Baltec Critical Point Dryer. | Once in 100% ethanol, samples are dried using a Baltec Critical Point Dryer. | ||
Revision as of 22:02, 27 October 2010
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Contents |
Scanning EM
This is a standard method for Scanning Electron Microscopy. Specimens vary greatly and the final protocol should be decided by referring to text books and publications.
Fixation
Fix in 2% Gluteraldehyde in Sorensons Phosphate Buffer overnight (minimum).
Rinse in Sorensons Phosphate Buffer 2x 15 mins.
Dehydration
- 25% ethanol: 30 mins.
- 50% ethanol: 30 mins.
- 75% ethanol: 30 mins.
- 100% ethanol: 1 hr.
- 100% ethanol: 1 hr.
Once in 100% ethanol, samples are dried using a Baltec Critical Point Dryer.
Mounting
Mount of an aluminium stub with Achesons Silver ElectroDag.
Gold coating
Coat with 15nm of gold using a Polaron SEM Coating Unit.
   
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