"
Team:Newcastle/Scanning EM
From 2010.igem.org
(Difference between revisions)
Swoodhouse (Talk | contribs) (→Dehydration) |
|||
| (2 intermediate revisions not shown) | |||
| Line 3: | Line 3: | ||
=Scanning EM= | =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. | + | *This is a standard method for preparing samples for Scanning Electron Microscopy. Specimens vary greatly and the final protocol should be decided by referring to text books and publications. |
==Fixation== | ==Fixation== | ||
| - | Fix in 2% Gluteraldehyde in Sorensons Phosphate Buffer overnight (minimum). | + | *Fix in 2% Gluteraldehyde in Sorensons Phosphate Buffer overnight (minimum). |
| - | + | *Rinse in Sorensons Phosphate Buffer 2x 15 mins. | |
| - | Rinse in Sorensons Phosphate Buffer 2x 15 mins. | + | |
==Dehydration== | ==Dehydration== | ||
| Line 16: | Line 15: | ||
*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. | + | |
==Mounting== | ==Mounting== | ||
| - | Mount | + | *Mount on an aluminium stub with Achesons Silver ElectroDag. |
==Gold coating== | ==Gold coating== | ||
| - | Coat with | + | *Coat with 15 nm of gold using a Polaron SEM Coating Unit. |
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Latest revision as of 22:14, 27 October 2010
| |||||||||||||
| |||||||||||||
Contents |
Scanning EM
- This is a standard method for preparing samples 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 on an aluminium stub with Achesons Silver ElectroDag.
Gold coating
- Coat with 15 nm of gold using a Polaron SEM Coating Unit.
   
|
    
|
