Journal of Applied Microscopy. 259 



8. Iron alum, a '1 per cent, aqueous solution of ammonio-sulphate of iron. 

 4. Hacmatoxylin, a 0.5 per cent, solution obtained by dissolving in hot 

 water. 



The remaining Stender dishes will therefore be as follows : 



1(1. Filled with anilin safranin. 



11. Filled with gentian violet. 



12. Filled with iron alum. 

 18. Filled with haematoxylin. 



First Stain — -Anilin Safranin. 



1. Melt the paraffin around the sections of two slides by heating them to 

 WlP C. in the paraffin oven. 



2. Wash off the paraffin by putting the two slides back to back into the 

 Stender dish with the turpentine. 



8. Transfer to Stender dish of xylol. 



4. Next put them in succession into the dishes with absolute alcohol, 95 per 

 cent, 85 per cent., 70 per cent., and 50 per cent. Let them remain in each one 

 about ten seconds, more or less. Do not leave the dishes uncovered longer than 

 necessary. 



5. Transfer the slides from the -50 per cent, alcohol to the anilin safranin 

 dish, and let them stain from two to four hours. 



Note. — In taking sections from xylol to any stain, always pass down the 

 grades of alcohol until the sections are in the same grade as the stain, and then 

 transfer to the stain. 



6. When the sections are stained wash them successively in the 50 per cent, 

 alcohol, TO per cent., 85 per cent., 95 per cent, and absolute alcohol. Judg- 

 ment must be used as to how fast the transfer is to be made from one grade of 

 alcohol to the other. They must generally be taken quite rapidly, as the alcohol 

 will take out such stains as safranin. 



7. Clear the sections by transfering them to the xylol. The sections' must 

 be thoroughly cleared. Leave them in xylol until they look transparent. 



8. Take one slide out of the xylol at a time ; wipe off the xylol with a clean 

 rag, wiping quite close to the sections, but do not touch the sections. 



9. .Put a drop or so of Canada balsam .(dissolved in xylol) on the sections at 

 one side. 



10. Put on a clean cover-glass in the following manner : holding the cover- 

 glass with the edges between the thumb and forefinger, bring it down slowly and 

 obliquely upon the drop so that one edge of it is first wetted by the balsam ; and 

 supporting the opposite edge with a needle, let the cover gradually settle down 

 and spread out the balsam. There should be no air bubbles and just enough 

 balsam to come to the edge of the cover-glass. Care must be taken to not let 

 the sections become dry at any stage of the foregoing process. The slides may 

 now be laid aside into a convenient place to dry. They may be studied imme- 

 diately if handled with care for a few weeks until the balsam has thoroughly 

 hardened around the cover-glass. If any balsam should get on the hands or 

 instruments, it can easily be removed with a little xylol. 



