Team:Newcastle/Scanning EM

From 2010.igem.org

(Difference between revisions)
(New page: {{Team:Newcastle/mainbanner}} =Scanning EM== This is a standard method for Scanning Electro Microscopy. Specimens vary greatly and the final protocol should be decided by referring to te...)
 
(6 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
-
=Scanning EM==
+
=Scanning EM=
-
This is a standard method for Scanning Electro 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==
-
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
+
==Mounting==
 +
*Mount on an aluminium stub with Achesons Silver ElectroDag.
 +
 
 +
==Gold coating==
 +
*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

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

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.
Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon