Journal of Applied Microscopy. 



135 



enough of this haematoxylin solution to 

 cover them. After this staining the pre- 

 parations are waslied for a few moments 

 in water, most conveniently done by 

 taking up each cover-glass with for- 

 ceps and holding it under flowing wa- 

 ter, or, if this is not at hand, by mov- 

 ing the preparation around in a relative- 

 ly large quantity of water. The prepara- 

 tions are now stained for about five 

 minutes in a one per cent, solution of 

 eosine, they are then again washed in 

 water, and dried between filter paper. 

 In order to insure perfect dryness, it is 

 well to hold the preparation for a few 

 minutes over a flame or radiator; they 

 may then be mounted on a drop of 

 Canada balsam. 



In blood preparations fixed and stained 

 after this method, the nuclei of all the 

 white blood cells are stained blue, also 

 of the red blood cells or of other ele- 

 ments if such are present. The red blood 

 cells are stained a light red by the 

 <:osine. 



The protoplasm of the white blood 

 cells, if stained at all, is a very faint 

 red. The only granules stained in pre- 

 parations fixed and stained after this 

 method, or the eosinophilous granules 

 of some of the white blood cells, and 

 these a bright red. 



EHRLICH'S NEUTROPHILE MIXTURE. 



One of the best staining solutions for 

 coloring blood preparations Is that sug- 

 gested by Ehrlich and known as the 

 "neutrophile mixture." It is somewhat 

 diflficult to prepare. The lollowing for- 

 mula is practically the same as that 

 used by Ehrlich: 



Orange G (8 parts to 100 water), 130 

 ccm. 



Acid Fuschin (20 parts to 100 water), 

 120ccm. 



Methylen Green (12 parts to 100 water), 

 125 ccm. 

 ' Distilled water, SOOccm. 



Absolute alcohol, 200ccm. 



Glycerine, lOOccm. 



Mix orange G, acid fuchsin, water, and 

 absolute alcohol in a bottle, add slowly 

 and while shaking the methylen green. 

 The glycerine is then added. For stain- 

 ing in Ehrlich's neutrophile mixture, the 

 preparations need to be hardened in 

 ether and alcohol for about one hour, or 

 fixed at a temperature of 100° to 110° C. 

 for fifteen to thirty minutes. Float the 

 preparation on a small quantity of the 

 stain for about fifteen minutes, wash in 

 flowing water, dry between several filter 

 papers, and mount in balsam. The red 

 corpuscles should have a. reddish brown 

 color (brick color), all nuclei green, the 

 eosinophilous granules red, and the 



neutrophile granules violet-red. The 

 granules found in the myelocytes, cells 

 found normally in the bone marrow, 

 and found in the circulation only in 

 pathological conditions, are stained like 

 the neutrophile granules a violet-red. 



It should be stated that, in order to 

 see the finer details in preparations 

 stained after either of the above 

 methods, it is necessary to employ a 

 one-tenth or one-twelfth-inch oil-immer- 

 sion lens. Without this aid it becomes 

 somewhat difficult to see the granulation 

 in the white blood cells. 



Histological Laboratory, University of 

 Michigan. 



( To be continued. ) 



Mounting Licliens. 



In the June number of the Journal, 

 Prof. George Pierce gives his method of 

 fixing and imbedding specimens of this 

 interesting group of plants. In the work 

 with my students I have pursued a dif- 

 ferent method from that given by Prof. 

 Pierce, and, as it has given good result^-, 

 I will give it for the benefit of the read- 

 ers of the Journal. 



• First the lichen is put into 95 per cent, 

 alcohol for 24 hours, then into thin cel- 

 loldin and thick celloidin for 24 hours 

 each. After this the specimens are im- 

 bedded in thick celloidin which is hard- 

 ened in 70 per cent, alcohol for 24 hours, 

 and then cut. I stain cryptogams gen- 

 erally with borax carmine as it gives 

 me better results than any other stain. 



In this case a medium staining with 

 borax carmine gives the fungus part of 

 the lichen pale carmine, while the pra- 

 tococcus cells have a greenish red shade. 

 This readily differentiates every cell of 

 the host from the fungus. 



Last week my class prepared sections 

 of two of our lichens in this way — Clad- 

 onia cornucopioides and Peltigera can- 

 ina — with excellent results. Other genera 

 have been treated in the same way by 

 former classes with equally good results. 

 G. H. French 



Department Biology and Physiology, 

 South 111. Normal University. 



An injecting mass suitable for gross 

 dissections can be made by stirring white 

 zinc, prepared chalk, or starch in linseed 

 oil varnish, to the consistency of soft 

 putty and again reducing to that of syrup 

 with ether. The mass can be colored by 

 stirring in chrome yellow, vermilion, or 

 other coloring materials. The mass 

 should be put into the vessels under as 

 heavy pressure as is possible with the 

 tissues treated. As the ether evaporates 

 the mass becomes hard. 



