DS 23.32 DYE STAINS OF SPECIAL APPLICATION 503 



preparation: Dissolve orange in solvents. Add violet, 



method: [fresh scrapings]-^ water, on slide -^ dry — > stain, on slide, 2 mins. — > 95% 



ale, 15 sees. — » water, 15 sees. — > dry 

 RECOMMENDED FOR: staining fungi in tissue scrapings. 



23.32 Bardelli and Cille 1928 test. 1942 Langeron Langeron 1942, 1251 



REAGENTS REQUIRED: A. acetic acid; B. DS 11.43 Ziehl 1882; C. ADS 12.2 Lugol; D. 



aniline 

 method: [thin sections] —> A, on slide, warmed gently, 30 sees.—* 96% ale, thorough 



wash — > dry — > B, 3 mins. — > drain or blot — > C, 5 mins. — > D, till differentiated — » 



balsam, via carbol-xylene 

 RECOMMENDED FOR: demonstration of Zymonema in sections. 



23.32 Berberian 1937 1750, 36:1171 



REAGENTS REQUIRED: A. 50% acctic acid; B. ether; C. acetone; D. DS 11.44 Martinotti 



1910 75, glycerol 20, 95% ale. 5; E. 0.5% acetic acid 

 method: [scrapings] -+ A, on slide — > dry -^ B, on slide, 20-30 sees. — »• C, 2 changes, 1 



min. each -^ abs. ale. — > [series ales, to 70%] — > D, 10-15 mins. -^ E, till differentiated 



-^ balsam, via acetone and xylene 

 recommended for: fungus hyphae in skin scrapings. 



23.32 Besson 1904 Besson 1904, 70 



REAGExNTs REQUIRED: A. DS 11.24 Orth (1924); B. DS 23.211 Ehrlich 1882; C. 0.7% 



sodium chloride; D. ADS 12.2 Gram 1884; E. anUine 

 method: [sections] ^ a, overnight -^ rinse -^ B, 20 mins. -^ C, rinse -^ D, some 



minutes -^ E, on slide, till differentiated — > balsam, via xylene 

 RECOMMENDED FOR: demonstration of Aspergillus fumigatus in sections of lung. 



23.32 Bernhardt 1943 1829,48:533 



REAGENTS REQUIRED: A. 10% potassium hydroxide; B. M 13.1 Bernhardt 1943 100, 



cotton blue 0.5; C. M 13.1 Bernhardt 1943 

 method: [skin scrapings or nail fragments] — > A, under cover, till clear and soft -^ press 



cover with blotter — > B, drawn under cover, warmed — > press cover with blotter -^ 



C, drawn under cover — »• seal 



23.32 Bigot 1924 5310, 17:547 



REAGENTS REQUIRED: A. DS 12.15 Crystal violet; B. ADS 12.2 Gram 1884; C 1% 



erythrosin 

 method: [smears, fixed in F 5000.1010 Duboscq and Brazil 1905 and depicrated in 



lithium carbonate sol.] — > A, 6-12 hrs. — » quick rinse — > B, 1 min. — > water, thorough 



wash — > C, 30 sees. — » water, wash -* dry 

 RECOMMENDED FOR: demonstration of Zymonema in tissue fragments or exudates. 



23.32 Boeck test. 1896 Kahlden and Laurent Kahlden and Laurent 1896, 146 



REAGENTS REQUIRED: A. DS 11.44 SahU 1885; B. 0.1% resorcin; C. 3% dilution of 30% 



hydrogen peroxide 

 method: [scales degreased in ale. and ether] -^ water — > A, 1-2 mins. — > B, J^ to 1 min. 



—>■ C, till differentiated — > wash — > balsam, via usual reagents 

 recommended for: fungi in skin scrapings. 



23.32 Chalmers and Marshall 1914 11587, 17:256 



reagents required: A. 40% potassium hydroxide; B. DS 23.211 Ehrlich 1882; C. ADS 

 12.2 Gram 1884; D. anihne; E. 2% eosin Y in 95% ale. 



method: [scrapings]—* A, some hrs., 40°C. — > 15% ale. thorough wash-^ strand on 

 slide -^ dry — > B, on slide, 20 mins. — > drain — > C, 3 mins. — > D, till no more color 

 comes away -^ E, 1 min. — ^ D, till no more color comes away — > balsam, via xylene 



recommended for: staining fungi in tissue scrapings. 



note: a detailed description of the use of this technique is given under DS 23.30 above. 



23.32 Curtis test. 1904 Besson Besson 1904, 699 



reagents required: A. DS 11.28 von Orth 1892; B. water 100, potassium hydroxide 

 0.01, methyl violet 6B 1.5; C. 1% pyrogallic acid 



