STAINING SPORES. 37 
stained, place again in the fuchsin. If the rods 
are still stained, wash longer in the acid alcohol. 
(/) Dry and pass.three times through the flame. 
(g) Stain in Léffler’s methylene-blue! for three 
to four minutes, or in gentian-violet? one minute, 
or in saturated aqueous solution of methylene-blue 
for half a minute. 
(2) Wash, dry, and mount. 
Spores should be red, rods blue. 
(2) Second Method. 
(a) Prepare films in the usual manner. 
(4) Place in chloroform for two minutes. 
(c) Treat with 5 per cent. chromic acid for one 
minute. The acid acts on the membranes of the 
spores and permits the entrance of the stain. 
(2) Wash thoroughly in water. 
(e) Stain in hot Ziehl’s carbol-fuchsin for three 
minutes. 
(f) Without washing decolorize in 5 per cent. 
sulphuric acid. 
(g) Wash in water and stain in Loffler’s methy- 
lene-blue' for two minutes or longer. 
(2) Wash, dry, and mount. 
(3) Third Method. 
(a) Suspend the spore-bearing bacteria in a nor- 
mal salt solution® in a test-tube. 
(6) Add an equal volume of Ziehl’s carbol-fuchsin. 
(c) Place the test-tube in boiling water for fifteen 
minutes. 
1 See Appendix, page 171. 2 See Appendix, page 171. 
3 See Appendix, page 173. 
