186 



HYPOPHYSIS AND GOXADOTROPHIC HORMONES 



tion of kre^ofuchsin and Heidenhain's azan 

 which, although differing in details from 

 the method of Berblinger and Burgdorf, 

 should have given, as far as the basophils 

 were concerned, equivalent results. His re- 

 sults are published in his magnificent mono- 

 graph (Romeis, 1940) on the hypophysis. 

 Unlike the results of Berblinger and Burg- 

 dorf, kresofuchsin in Romcis's hands gave a 

 clear-cut discrimination between two dis- 

 tinctive groups of cells, those whose gran- 

 ules stained with kresofuchsin and those 

 whose granules stained not with kresofuch- 

 sin but only with the aniline blue of the 

 azan stain. During his investigation Romeis 

 found that the kresofuchsin supplied by the 

 manufacturers differed from that he had ob- 

 tained earlier in that, while still staining 

 elastic tissue, it was no longer useful for 

 staining the hypophysis. He found that 

 resorcin-fuchsin he made himself could be 

 substituted for kresofuchsin with results 

 which were equal in every respect. 



Romeis examined the hypophyses of a 

 number of mammalian species other than 

 man with results that differed from those 

 of Rodriguez and von Soos in some respects. 

 He found cells with granules stainable by 

 kresofuchsin or resorcin-fuchsin in some 

 species in which these authors had not been 

 able to demonstrate them. As has been 

 stated earlier, Romeis called the cells whose 

 granules stained with kresofuchsin ^-cells, 

 and those whose granules did not stain with 

 kresofuchsin, but were stained with aniline 

 blue, 8-cells. He found both /?- and 8-cells 

 in the bovine hypophysis and in the hy- 

 pophyses of the dog, horse, cat, and rat. He 

 found /3-cells alone in the bat and guinea 

 pig; in the latter they were very scanty. He 

 also noted that kresofuchsin stained strongly 

 the pars intermedia cells in the guinea pig, 

 mouse, and pig. 



The results that Romeis obtained with 

 resorcin-fuchsin have been duplicated by 

 Ezrin, Swanson, Humphrey, Dawson and 

 AVilson (1958). Despite many attempts, 

 other investigators have not been able to 

 repeat these results using Romcis's method. 

 There is, however, no doubt about the cor- 

 rectness of his observations. 



Gomori (1950) introduecul ahU^liydo- 

 fuclisin as an elastic tissue stain, and Ilahni 



(1950, 1951a) used it to differentiate be- 

 tween ^- and 8-cells in the rat and mouse. 

 It has since been used successfully by a 

 number of authors and on a number of spe- 

 cies. The question whether aldehyde-fuchsin 

 gives results equivalent to those once given 

 by kresofuchsin is one which is not easily 

 answered. The results are variable, depend- 

 ing on the fixative and on the treatment of 

 the slide before staining, so that the term 

 "stainable by aldehyde-fuchsin" cannot be 

 given any precise meaning. Moreover, there 

 are in the pars anterior of many, if not all 

 mammals, three types of basophil cells, and 

 according to conditions, the aldehyde-fuch- 

 sin may stain the granules of one, two or 

 all three types, or fail to stain any of them. 

 I think, however, that it may be assumed, 

 that with any particular fixative and prior 

 treatment of the slide, the elements most 

 readily stained by kresofuchsin or resorcin- 

 fuchsin would also be most readily stained 

 by aldehyde-fuchsin. 



Aldehyde-fuchsin suffers from the defect 

 of variability in staining power from batch 

 to batch, and it often happens, as Romeis 

 found with kresofuchsin, that a batch of 

 this stain will stain elastic fibers but fail to 

 stain basophil granules. Gabe (1953) de- 

 scribed the preparation of an aldehyde- 

 fuchsin which can be kept as a dry powder. 

 To obtain results equivalent to those given 

 by Gomori's preparation it is necessary to 

 dissolve this powder in a concentration of 

 0.5 per cent in 70 per cent alcohol acidified 

 with hydrochloric acid. Gabe's own proce- 

 dure does not give the same results. 



Recently Herlant and Racadot (1957) 

 have indicated that Alcian blue (Steedman, 

 1950) at pH 1 stains the basophil granules 

 in the toad and cat hypophysis that are 

 stained by aldehyde-fuchsin. I find that a 

 3 per cent solution of Alcian blue in 70 per 

 cent alcohol acidified with hydrochloric 

 acid stains rat and human hypophyses with 

 results equivalent to those obtained with 

 aldehyde-fuchsin except that elastic fibers 

 are not stained. H this procedure proves 

 equally useful in other species it may sup- 

 plant aldehyde-fuchsin and thus eliminate 

 the uncertainty attending the use of this at 

 |)resc>nt indispensable stain. 



