Team:Newcastle/Scanning EM

From 2010.igem.org

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(Fixation)
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==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.

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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