DS 23.214-DS 23.215 DYE STAINS OF SPECIAL APPLICATION 481 



23.214 Tunnicliff 1921 11006, 78:191 



REAGENTS REQUIRED: A. DS 12.15 Crystal violet; B. ADS 12.2 Lugol 1905; C 1% 



safranin O 

 method: [heat-fixed smear] -^ A, 30 sees. -^ water, wash — > B, 30|sccs. — > water, wash -> 

 C, 30 sees. — » water, wash — > dry 



23.214 Weiss 1929 11284, 14:1191 



REAGENTS REQUIRED: A. acetic acid ; B. ADS 12.2 Weiss 1929; C. sat. aq. sol. crystal 

 violet or safranin or magenta or brilliant green; D. 10% acid green or acid violet or 

 acid fuchsin in 70% ale. 

 method: [drop material and drop A incubated as hanging drop, 37.5°, 15 mins.] -+ air- 

 dried smear — » B, on slide, heated to steaming, 2-5 mins. -+ wash — > C, 2-5 mins. -^ 

 wash — » D, 8-10 mins. -^ wash — > dry 

 note: The selected dyes in C and D above should be of contrasting colors. 



23.214 Woolman 1939 test. 1939 Findlay 11360, 59:184 



REAGENTS required: A. watcr 97, 40% formaldehyde 2, acetic acid 1; B. water 100 



hydrochloric acid 0.7, crystal violet 0.15, copper sulfate 10 

 method: [air-dried smears]-^ A, 3-5 mins. -+ wash — > abs. ale, 1-2 mins. — > blot—* 



B, 10-15 mins. — > drain — * dry 



23.215 Flagella Stains 



The term flagella stains is somewhat misleading because there is no known technique 

 which could be employed differentially to stain a flagella protruding from a bacteria 

 without staining every other minute particle of the protein material present. The pur- 

 pose of these stains, however, is to provide so dense a deposition of dye as to assure 

 that these minute particles will become so covered as to be visible in an optical micro- 

 scope. For this reason every possible type of mordant, usually if not always combining 

 tannin with iron, is used before a solution either of one of the thiazins or of magenta. 

 The real difficulty in the staining of flagella is not to deposit the stain, but to differ- 

 entiate the subsequently stained flagella from the background. For this reason the very 

 utmost care must be taken in the cleanliness of the slides employed for the preparation 

 of a smear, and all those precautions proper to metal stains (which are given in Chap- 

 ter 24) can with justice be used in the present instance. If, however, the flagellated 

 organisms are taken from a culture filled with small particles of organic detritus, it is 

 a waste of time to endeavor to stain the flagella on them. A recent method of Fisher 

 and Conn 1942 gives the most minute directions for the selection for the culture as well 

 as for staining the organisms. 



23.215 Bailey 1929 16913, 27:111 



reagents required: A. ADS 12.2 Bailey 1929; B. ADS 12.2 Bailey 1929 70, DS 11.43 

 Ziehl 1882 10, hydrochloric acid 10, 40% formaldehyde 10; C. DS 11.43 Ziehl 1882 



method: [dry smear] -^ A, 2 mins. — > 5, 7 mins. — > wash — > C, warmed to steaming, 30 

 sees. — * wash -^ dry 



23.215 Bowhill 1898 23684, 23 :667 



preparation of stock soiitrriONs: I. water 40, 95% ale. 50, orcein 1; II. water 100, 



tannin 20 

 reagents required: A. stock I 50, stock II 50; B. DS 23.211 Ehrlich 1882 

 method: [heat-fixed smear] — > A, 10 mins. at 50°C. — * wash -^ dry -* B, heat to steam- 

 ing, 30 sees. — * wash -^ dry 



23.215 Bunge 1894 8645, 12 :24 



formula: water 100, tannic acid 100, ferric chloride 1.4, magenta 0.25, hydrogen per- 

 oxide q.s. 



preparation: Dissolve the tannic acid and ferric chloride in 90 water. Add the dye dis- 

 solved in 10 water. Add enough hydrogen peroxide just to turn solution brown. 



method: As DS 23.215 Loffler 1890 



