Post Staining
- Place 1 drop of stain for each grid on PARAFILM.
- Place grid on top of stain drop, section side down.
- Cover with Petri dish lid to avoid dust and evaporation.
- Stain for 30 minutes.
- Set up four 50 ml. beakers of Glass Distilled Water.
Rinse grids in first beaker 4 or 5 dips to remove most of stain.
Then, dip each grid in remaining 3 beakers, 30 dips each.
Touch to Whatman 1 filter paper to drain off water.
Let dry a couple of minutes before proceeding to Lead stain.
LEAD CITRATE STAINING (0.3% Lead Citrate in GD Water, pH 12.0)
- Proceed as in Steps 1 - 3 above.
- Stain for 5 minutes. If you stain more than one grid, then give yourself about a 2-minute interval between the start of each grid to allow time for washing.
- Wash each grids 30 dips in 3 beakers of GD Water.
- Touch to Whatman 1 filter paper to drain off water.
NOTES
- After staining, as you transfer grids to Petri dish for storage, gentley push grid off forceps with filter paper to remove water at forcep tip. Otherwise, capillary action will suck the grid between the points and perhaps damage sections.
- Keep grids covered during staining, especially during lead stain, to avoid lead carbonate crystals from forming from exposure to room air.
- When handling bottles of stain, do it gently. Avoid stirring up the contents of bottles.
- Place PARAFILM containing stain and pasteur pipets in hazardous waste container in exhaust hood.