Determination of Altered Mitochondria Ultrastructure by Electron Microscopy - Protein Misfolding and Cellular Stress in Disease and Aging - page 274

(b) Stain the sections with 0.5% toluidine blue for several
minutes at 150-200°C.
(c) Wash the slide glass in tap water or distilled water and
then dry it on the heater, and if necessary, mount the
sample.
19. The interference color of the section serves as an index of its
approximate thickness: gray, 50-60 nm; silver, 70-90 nm;
and gold, 90-150 nm. The author usually prefers silver-gold
or gold coloring for tissues of the nervous system. The ultra-
thin sections are collected onto grids. From among a variety of
grids from coarse-meshed to fine-meshed, the present author
prefers a grid of 150 meshes from the practical point of view.
20. Stain the sample with uranyl acetate for 5 min (Fig. 1a ),
followed by a 50% ethanol wash and a wash in distilled water
(Fig. 1c ). Then, stain the sample with lead citrate for 2 min
(Fig. 1b ), followed by a wash in distilled water (Fig. 1c ) and
subsequent drying. To create the uranyl acetate staining
Fig. 1. Sample staining. For explanation see Note 20. ( a ) Place the side of the grid on
which the tissue is attached on the surface of the uranyl acetate solution. ( b ) Sink the
water washed grid with the tissue attached to the bottom of the lead citrate solution.
( c ) Prepare two bottles for washing the grid with the tissue attached. One of the bottles
should contain a 50%-ethanol solution, and the other should contain distilled water. Pick
up the uranyl acetate-stained grid with tweezers, and wash it directly with 50%-ethanol
solution. Pick up the lead citrate-stained grid with tweezers and wash it directly with the
distilled water.