SIMPLE METHODS FOR “STAINING WITH LEAD” AT HIGH pH IN ELECTRON MICROSCOPY

SIMPLE METHODS FOR “STAINING WITH LEAD” AT HIGH pH IN ELECTRON MICROSCOPY

July 24, 1961 | MORRIS J. KARNOVSKY.
This article presents two methods for preparing highly alkaline lead staining solutions for electron microscopy, which are simpler and more stable than previous methods. The first method involves dissolving lead monoxide in hot NaOH to produce the plumbite ion, while the second method uses lead monoxide dissolved in sodium cacodylate followed by alkalization with NaOH. Both methods yield solutions with a pH > 11.5, which are stable and produce rapid, intense staining without the precipitation issues of lower pH solutions. The staining procedure is described in detail, including the use of a staining chamber and the optimal staining times for different materials. The results show that the staining intensity is comparable to that of Watson's lead hydroxide technique, with minor differences. The mechanism of staining is discussed, suggesting that the plumbite ion is responsible for the staining, possibly through hydrogen bonding to tissue groups. The article also highlights the advantages of the methods, such as ease of preparation, stability, and the ability to stain difficult materials.This article presents two methods for preparing highly alkaline lead staining solutions for electron microscopy, which are simpler and more stable than previous methods. The first method involves dissolving lead monoxide in hot NaOH to produce the plumbite ion, while the second method uses lead monoxide dissolved in sodium cacodylate followed by alkalization with NaOH. Both methods yield solutions with a pH > 11.5, which are stable and produce rapid, intense staining without the precipitation issues of lower pH solutions. The staining procedure is described in detail, including the use of a staining chamber and the optimal staining times for different materials. The results show that the staining intensity is comparable to that of Watson's lead hydroxide technique, with minor differences. The mechanism of staining is discussed, suggesting that the plumbite ion is responsible for the staining, possibly through hydrogen bonding to tissue groups. The article also highlights the advantages of the methods, such as ease of preparation, stability, and the ability to stain difficult materials.
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