BUNQE'S METHOD. 157 



be said that bacteria which produce acids in the media 

 in which they are,grovving require the addition of alka- 

 lies to the mordant, while those that produce alkalies 

 require acids to be added. By following LoelHer's direc- 

 tions the delicate, hair-like flagella on motile organisms 

 may be rendered plainly visible. 



There are several points and slight modifications in 

 connection with this method that require to be empha- 

 sized in order to insure success: the culture to be em- 

 ployed should be young, not over 18-20 hours old. It 

 should have developed for this time on fresh agar-agar 

 at 37° to 38° C; the mordant should not be perfectly 

 fresh, as the best results are obtained from the use of 

 old solutions that have stood exposed to the air, and 

 that have been filtered just before using; when placed 

 on the cover-slip and held over the flame never heal the 

 mordant to the boiling-point ; indeed, the best results are 

 obtained when the preparation is held high above the flame 

 and reftnoved from it at the first evidence of vaporization, 

 or, better still, a little before this point is reached. AVe 

 have deri\'ed no advantage from the addition of acids 

 or alkalies to the mordant, as recommended by Loeffler; 

 but obtain, with a fair degree of regularity, satisfactory 

 results through the use of the neutral mordant alone.^ 



Bunge's Method. — A useful modification of Lcef- 

 fler's method is that recommended by Bunge: prepare 

 a saturated solution of tannin, and a solution of liquor 

 ferri sesquichlor. of the strength of 1 : 20 of distilled 

 water. To 3 parts of the tannin solution add 1 part of 

 the dilute iron solution. To 10 c.c. of such a mixture 



1 I am indebted to Dr. James Homer Wright, Thomas Scott Fellow in Hy- 

 giene, 1892-93, University of Pennsylvania, for some of the suggestions in 

 connection with the modification of this method. 



