Team:Newcastle/Scanning EM

From 2010.igem.org

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=Scanning EM=
=Scanning EM=
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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.
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*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==
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Fix in 2% Gluteraldehyde in Sorensons Phosphate Buffer overnight (minimum).
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*Fix in 2% Gluteraldehyde in Sorensons Phosphate Buffer overnight (minimum).
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*Rinse in Sorensons Phosphate Buffer 2x 15 mins.
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Rinse in Sorensons Phosphate Buffer 2x 15 mins.
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==Dehydration==
==Dehydration==
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*100% ethanol: 1 hr.
*100% ethanol: 1 hr.
*100% ethanol: 1 hr.
*100% ethanol: 1 hr.
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*Once in 100% ethanol, samples are dried using a Baltec Critical Point Dryer.
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Once in 100% ethanol, samples are dried using a Baltec Critical Point Dryer.
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==Mounting==
==Mounting==
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Mount of an aluminium stub with Achesons Silver ElectroDag.
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*Mount on an aluminium stub with Achesons Silver ElectroDag.
==Gold coating==
==Gold coating==
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Coat with 15nm of gold using a Polaron SEM Coating Unit.
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*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

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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.
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