UranyLess – Protocols of Use: Classic Contrast

EMS Catalog #22409

This protocol is used for double staining with UranyLess / Lead citrate on ultrafine sections.

This protocol is adapted to biologic samples that have been fixed with glutaraldehyde, osmium, or ruthenium and embedded in an epoxy type resin (Epon, Araldite, Spurr) or acrylic type (LRWhite, HM20).

Staining protocol:

1. Place a drop of UranyLess on parafilm or any other hydrophobic slide.
2. Place the grid on the UranyLess drop for 1 to 2 minutes.
3. Blot the grid on a filter paper and then wash in room temperature distilled water.

Let it dry.

After drying, go to the lead citrate staining according to Reynolds method (1963).

1. Place the grid on the lead citrate drop according to the Reynolds method, for 1 minute.
2. Blot the grid on a filter paper before rinsing with room temperature distilled water.
3. Let it dry.

Technical Tip:

• UranyLess is not air or light sensitive, unlike Uranyl Acetate.
• After lead citrate, drain immediately in a freshly prepared room temperature distilled water bath or wash with 0.01N of NaOH solution. Cold water is charged with CO₂ and causes precipitate on the grid.
• If there is a precipitate in the solution, filter it prior to use.
• If solution was refrigerated, allow solution to return to room temperature prior to use.
• Do not keep lead citrate refrigerated.

Product Information

UranyLess, Substitute for Uranyl Acetate