Silver Enhancement Procedure
using AURION R-Gent SE-EM
(Silver Enhancement for Electron Microscopy)
It is assumed that the Developer has been prepared as prescribed.
1. Prepare the enhancement mixture as
follows:
Once the DEVELOPER and ENHANCER have reached room temperature, give
20 drops of the ENHANCER solution into a vial that will contain at least 1.5 ml,
e.g. an Eppendorf vial. Make sure to keep the bottle upside down in a vertical
position. Add 2 drops of the DEVELOPER solution, again making sure that
the bottle is kept upside down in a vertical position. Mix well on a
vortex.
2. Specimens are floated in enhancement mixture on a rocking table. Enhancement time is typically between 30 minutes and 1 hour at room temperature (preferably 20°C). The actual enhancement time has to be established empirically and adjusted according to the desired particle growth.
3. When enhancement is complete the specimens are washed extensively with distilled water (at least 3x10 minutes). A postfixation with photographic fixer is not required.
4. Specimens are dehydrated and embedded according to standard procedures.
Silver enhancement and OsO4
fixation
Silver enhancement may be applied either before or after
OsO4 fixation.
The time of enhancement may have to be longer due to potential removal of silver by OsO4.
The fixation step is inserted before dehydration.
Enhancement after OsO4 fixation
The fixation step is inserted
before silver enhancement.
Wash extensively after OsO4 fixation: traces
of this strong oxidizing agent will interfere with adequate silver
enhancement.
Alternatively enhancement may be done after ultra thin
sectioning.
RECOMMENDED FOR EM
- Incubation
Solution
- AURION Blocking Solutions
- AURION BSA-c
- AURION Ultra Small Immuno Gold Reagents
- AURION Conventional Reagents
(when using Ultra Small IGR)
For post-embedding
Glycine to prepare the aldeyde blocking solution
For pre-embedding
Sodium Borohydride to prepare the aldehyde blocking solution
Triton-X-100 for increased accessibility
