Stains, Reagents, Materials:
Saturated uranyl acetate in 75% methanol
Lead citrate – from a mixture of Na citrate and Pb nitrate
KOH pellets – for mopping up CO2
Whatman filter paper #1
Parafilm
Al foil for a clean work area
Glass Petri dishes
6 rinse bottles (scint vials)

Protocol:

• Fill rinsing bottles with dH2O
• Filter UA solution into clean scintillation vial
• Put KOH pellets at edge of Petri
• Place drops of UA on parafilm (one per grid)
• Float grids, section side down on UA drops for 6 minutes
•     Stagger the grids by 2 minutes
•     Keep track of which is which!
• Transfer to rinse #1 (don’t let them dry in between)
•     Jiggle up and down in rinse water
•     Then rinse with dH2O from squirt bottle (not directly)
• Repeat with rinses 2,3
• Transfer grid to filter paper using filter paper wedge
• Let dry completely
• Rinse and clean forceps before starting Pb citrate
• Handle bottle of Pb citrate carefully to avoid precipitation
• Repeat with Pb citrate, but only stain for 4 minutes