A double lead stain method for enhancing contrast of ultrathin sections in electron microscopy: a modified multiple staining technique

Abstract

A modification of the conventional method for the staining of ultrathin sections resulted in an increase in contrast of ultrastructural detail in tissues. Tissues embedded in Spurr's low viscosity embedding medium were stained with freshly centrifuged Reynolds' lead citrate for 1-5 min, rinsed in double distilled water and dried prior to staining with a saturated solution of uranyl acetate for 40 min, and freshly centrifuged Reynolds' lead citrate for 20 min.Sections treated by this procedure showed enhanced staining of cellular organelles and cytoplasmic matrix. This procedure is recommended for tissues with poor staining qualities resulting from either prolonged fixation or from inadequacies in the buffer or embedding medium used.

Metformin Induces Ultrastructural Alterations in Hepatocytes of Spontaneously Hypertensive Rats

“…Silver-gold (60-90 nm thick) sections were selected and mounted on 300-meshed copper grids. Sections were then stained first with lead citrate for five minutes, then in saturated aqueous uranyl acetate (10%) for 40 minutes, then in freshly centrifuged or filtered lead citrate for another 20 minutes (Daddow, 1983).…”

An ultrastructural study of the spermatozoa ofEulalia sp. (Phyllodocidae),Lepidonotus sp. (Polynoidae),Lumbrineris sp. (Lumbrineridae) andOwenia fusiformis (Oweniidae)

“…Thin sections, 500-800 A thick, were collected on carbon stabilized colloidincoated 200 mesh copper grids. Sections were stained in either of two methods: (1) 40 min in a 10 % super-saturated solution of aqueous uranyl acetete, rinsed in distilled water followed by 20 min in Reynold's lead citrate and rinsed again before viewing; (2) a modification of the Daddow (1983) …”

Intracellular biogenesis of collagen fibrils in ‘activated fibroblasts’ of tendo Achillis

“…The samples were embedded in Araldite and Spurr resins. We obtained semithin sections which were stained with Toluidine Blue to identify the region of regeneration and ultrathin sections which were stained with lead citrate 84 or uranyl acetate 85 and observed under transmission electron microscopy.…”