DS 23.221-DS 23,222 DYE STAINS OF SPECIAL APPLICATION 493 



23.221 Mallory 1938 Mallory 1938, 86 



STOCK solutions: I. water ]()(), sodium borate I, mctliylcne blue 1; II. 1% azur TI 

 REAGENTS REQUIRED: A. 2.5% plilo.xiiie; B. stock I 5, stock 11 5, water 90; ('. ADS 21.2 



Wolbach 1911 

 method: a, 1-2 hrs., 50°C. -^ water, wash -> B, 5-20 inins. — > water, wash — > C, till 



differentiated — > balsam, via xylene 

 note: a detailed description of the use of this technique is given under DS 23.20 above. 



23.221 Masson see DS 13.22 Masson (1942) or DS 13.5 Masson (1942) 



23.221 Nicolle test. 1904 Besson Besson 1904, 258 



REAGENTS REQUIRED: A. J)S 11.44 Kiihnc (1904) a or b; B. 10% tannic acid 

 method: [sections] -^ water — > .1, 2 3 mins. -^ wash — > B, a few sees. —> rinse — ♦ blot — > 

 dehj'drate least possible time — ^ balsam, via usual reagents 



23.221 Nicolle see DS 11.423 Nicolle (1942) 



23.221 Noniewicz test. 1896 Kahlden and Laurent Kahlden and Laurent 189G, 108 



reagents required: A. DS 11.44 Loffler 1890; B. water 99, acetic acid 1, tropeolin 0.1 

 method: [sections] -^ water -^ A, 2-5 mins. -^ wash — > B, 1-5 sees. — > wash — > dry 



23.221 Ollett 1947 11431,69:357 



reagents required: A. DS 23.211 Ehrlich 1882; B. ADS 12.2 Gram 1884; C. 2% acetic 



acid in abs. ale; D. water 75, DS 13.7 Twort (1909) 25 

 method: [3 fj. sections of formaldehyde-fixed material]—* A, 3-5 mins. -^ rinse—* B, 



3 mins. -* rinse —* blot -^ C, till decolorized — > rinse — > D, 5 mins. —> rinse -^ C, till 



no more color comes away -^ balsam, via xylene 



23.221 Pappenheim DS 23.221 see Saathof 1905 



23.221 Saathof 1905 Unna-Pappenheim method — compl. script. 



7276, 32 :2047 

 formula: water 75, phenol 1.5, glycerol 20, 95% ale. 5, pyronin 0.15, methyl green 0.5 

 method: [sections] —* stain, 1-3 mins. — * w^ater, wash — > acetone, till dehydrated -^ bal- 

 sam, via xylene 



23.221 Unna see DS 23.221 Saathof 1905 



2S.222 Iodine Differential Methods 



Staining Gram-positive bacteria sections is only another instance in which a standard 

 method has become so embedded in the literature that all other methods are referred 

 to it. In this instance it is the Gram-Weigert technique which dominates the field. 

 Gram, in point of fact, had nothing to do with the technique of Weigert, published in 

 1887, which depended entirely upon Weigert's contribution of differentiating with 

 aniline the crystal violet stain of Ehrlich, after mordanting with Gram's iodine. So 

 confused, how^ever, has the picture become, that even Weigert's well-known resorcin- 

 magenta method for staining elastic fibers has been recommended for the purpose of 

 differentiating bacteria. Moreover, some authors (see comments under "Gram-Weigert" 

 below) have recommended prior nuclear staining either with carmine or hematoxylin. 

 Most of the techniques recommended here still rely on differentiation with aniline, 

 though the method of Brown and Brenn 1931 substitutes a mixture of acetone and 

 ether for the aniline. 



23.222 Brown and Brenn 1931a 10919, 48:69 



reagents required: A. DS 11.123 Harris 1900; S. 3% hydrochloric acid in 95% ale; C. 



1% ammonia; D. DS 11.45 Brown and Brenn 1931; E. ADS 12.2 Lugol; F. 0.005%, 



rosanilin hydrochloride; G. 0.1% picric acid 

 method: [sections]-^ water -^ A, 2-5 mins.—* rinses B, till pink—* rinse—* C, till 



blue — * wash — > Z), 2 mins. — * wash — > ^, 1 min. -^ wash — > blot — * F, 5 mins. — > 



wash -^ blot -^ acetone, rinse — > G, till yellowish pink -^ balsam, via acetone and 



xylene 

 result: Gram -positive organisms, red; Gram -negative organisms, blue black. 



