Difference: PPFiducialsTomography (13 vs. 14)

Revision 1419 Aug 2009 - Main.KdDerr

 
META TOPICPARENT name="CemProtocGrids"
Deleted:
<
<		 NYSBC|Cryo-EM P&P: Grid preparation - Fiducials for tomography
 
Changed:
<
<		Principles & Protocols
>
>

Principles & Protocols

Added:
>
>
 
Changed:
<
<
>
>

Application of Gold Beads to sections as Fiducial Markers for Tomography

Deleted:
<
<
Interested in EM? Using CEMfac? Principles & Protocols CEMfac Equipment Seminars & Courses
CEMfac Home? CEM Introduction Microscope Schedule NYSBC Directory Cryo-EM public website

Grid preparation

Protocols - Fiducials for tomography

Contents

Application of Gold Beads to sections as Fiducial Markers for Tomography
 Gold Fiducial markers can be applied before or after the sections are placed on the grid. If they are applied before the sections, a thin film must be present for the gold to adhere to.

You will need:

  • 200 mesh grids with sections or a thin film
  • Parafilm
  • Sonicator
  • Colloidal gold solution 5 & 10nm
  • DD H2O
  • Filter paper
  • Tweezers
  • glow discharge

Procedure:

  • Put on gloves!
  • Positive stain sample grids
  • Sonicate gold solution 10-15 min
  • Glow Discharge the grid with sections
  • Place grid sample side up on parafilm
  • Deposit 1um of 10 nm gold solution
  • Allow to air dry
  • Apply sample or view grid

Do not use copper grids for immunogold labeling

Public website mirror

-- DavidStokes - 18 Mar 2005

META TOPICMOVED by="KakoliMitra" date="1234475869" from="Main.CemPPGridsFTO" to="Main.PPFiducialsTomography"
View topic  |  History: r16 < r15 < r14 < r13  |  More topic actions...
 
Copyright © by the contributing authors. All material on this collaboration platform is the property of the contributing authors.
Ideas, requests, problems regarding this intranet, Send feedback