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Uranyl Acetate (UA) en bloc Protocol

This is used to bring out unit membranes.
The stain is used just after osmium post-fixation and before dehydration.
It usually dissolves glycogen or renders it unstainable.

Stocks

  • pH 5.15 Maleate buffer: 11.6 g maleate/500 ml dH2O, adjusted to pH 5.15 with NaOH
  • pH 6.00 Maleate buffer: 11.6 g maleate/500 ml dH2O, adjusted to pH 6.00 with NaOH
  • 1% uranyl acetate in pH 6.00 maleate (pH drops on addition of uranyl acetate; dissolve by sonication)

Procedure

  1. After osmication, wash tissue blocs in pH 5.15 maleate buffer (3 x 5 min).
    If phosphate buffer was used previously, more extensive rinsing will be
    necessary to prevent precipitation of uranyl phosphate. For thin retinal
    tissue, 3 x 15 min should suffice.
  2. Stain in 1% uranyl acetate in pH 6.00 maleate for 1 hr at room temperature wrapped in foil (i.e. dark).
  3. Wash in pH 5.15 maleate buffer (3 x 5 min).
  4. Proceed to dehydration.

REFERENCES:

Farquahar and Palade, J CEll Bio 26:263, 1965.

Trelstad, Revel and Hay, J Cell Bio c-6, 1966.


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