STAINING METHODS 



This gives a very sharp differentiation of bacteria and nuclear structures. It 

 has a purplish tinge. Fixation by heat gives the best staining. Allow the stain to 

 act for two to ten minutes. It should not be used until after standing twenty-four 

 hours, and after standing about two weeks it appears to lose its sharp staining power 



Archibald's Stain. This is an excellent bacterial stain and has been 

 highly recommended by Blue and McCoy in plague work. 



SOLUTION No. i. 



Thionin 0.5 



Phenol crys 2.5 



Formalin i.o 



Water 100.0 



SOLUTION No. 2. 



Methylene blue 0.5 



Phenol crys 2.5 



Formalin i.o 



Water 100.0 



Dissolve for twenty-four hours. Mix equal parts and filter. Stain smears fixed 

 by heat or otherwise for ten seconds. 



Nicolle's Carbol Thionin 



Sat. sol. thionin in 50% alcohol 



Carbolic acid solution (2%). 



10 c.c. 

 100 c.c. 



Pappenheim's Stain. Take a very small portion of methylene green on the point 

 of a penknife and shake it into a test-tube. Then take up twice as much pyronin and 

 deposit it in the same test-tube. Fill the test-tube one-half full with water and the 

 solution should have a distinct reddish-violet color. A drop on a piece of filter-paper 

 shows a violet center and peripheral green ring. The solution should be fresh. 

 Stain from two to five minutes. Differentiate with a little resorcin on a penknife 

 point dissolved in one-quarter of a test-tube full of alcohol. Dehydrate, clear and 

 mount. Polymorphonuclear nuclei stain greenish; nuclei of mononuclears and 

 plasma cells from bluish-red to dull violet. Cytoplasm of lymphocytes and plasma 

 cells purplish-red. Bacteria red. 



Romanowsky Stains. See under section on Blood. For mounting 

 specimens showing chromatin staining, as malarial parasites, trypano- 

 somes, intestinal flagellates, etc., liquid petrolatum is to be highly rec- 

 ommended. The chromatin staining lasts without any fading for at 

 least two years. The acidity of balsam causes rapid fading of the 

 chromatin. 



Neisser's Stain for Diphtheria Bacilli 

 SOLUTION No. i SOLUTION No. 2 

 Methylene blue. ... o. i gram. Bismark brown 



0.2 



Alcohol 2 c.c. 



Glacial acetic acid. . 5 c.c. 



Distilled water 95 c.c. 



Dissolve the methylene blue 

 in the alcohol and add it to 

 the acetic acid water mixture. 

 Filter. 



Water (boiling) 100 c.c. 



Dissolve the stain in the boil- 

 ing water and filter. 



