An abbreviated method of the double lead stain technique

An enhanced staining of cellular organelles and cytoplasmic matrix is observed when ultrathin sections for electron microscopy are stained for 1-5 min in fresh Reynolds' lead citrate, rinsed in distilled water and dried prior to staining with a saturated solution of uranyl acetate (40 min) and Reynolds' lead citrate (20 min), Daddow (1983). The above procedure was recommended for tissues with poor staining qualities either from prolonged fixation or inadequacies of the buffer or the embedding medium used. Using an abbreviated method of the above (i.e., 30 sec lead citrate, 60 sec saturated uranyl acetate and 30 sec lead citrate), overfixed or normally fixed tissues show enhanced contrast. Sodium hydroxide (0.02 M), substituted for the initial lead stain, increases contrast to a lesser extent.