By Geoffrey H. Bourne
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Moore and S. N. Ray, Nature 130, 997 (1932). 26 GEOFFREY H. BOURNE were not able to reduce it in the acid solution or would do so only after a long time in the light. The final technique established for the demonstration of this vitamin was, first, that scrupulous chemical cleanliness was required and, second, that impregnation should be for short periods of time (not more than 20 minutes) and should be carried out in the dark. After impregnation the tissues were treated with photographic "hypo" to remove silver compounds which might subsequently be reduced by the action of light.
11, 261 (1933). 102 G. H. Bourne, Nature 132, 859 (1934). 104 The principle of the technique is based upon the fact that the vitamin is a powerful reducing agent and that it will reduce silver nitrate not only in alkaline solution, as many other biological substancet do, but also in neutral and acid solution. The first preparations by Bourne100·101 were made with neutral silver nitrate. First attempts to demonstrate the vitamin were made on the adrenals (which are extremely rich in the vitamin) of cat and guinea pig which had been fixed in the vapor of formaldehyde.
Riboflavin in the convoluted tubule cells of the guinea pig kidney. The granules of riboflavin are similar in form and distribution to mitochondria. (Courtesy of D r . F . ) C. U. of vitamin A. Strong vitamin A fluorescence is shown by droplets within the lumen, and moderate fluorescence by the epithelial lining and by cells in the center of the villi. (Courtesy of D r . ) D. Thiamine (vitamin Bi) in the sciatic nerve of the rabbit. T h e reaction is probably due to the spontaneous oxidation of thiamine to thiochrome.
Biochemistry And Physiology of Nutrition, Vol. 2 by Geoffrey H. Bourne