471 



NOTE ON A QUICK METHOD OF PREPARING AND 



STAINING POLLEN. 



By W. Wesche, F.E.M.S. 

 {Read October 2Qth, 1909.) 



After trying staining the grains and clearing them in phenol 

 and xylol, which failed to remove the stain and also presented 

 great difficulties in manipulation, I tried the following quick 

 method, which answered admirably. 



The flowers were collected during the period August lst-15th ; 

 they were kept in pill boxes till October 2nd, when the experi- 

 ment was made.* They were shaken on to a slip and scraped 

 with a needle to free the pollen ; the debris other than pollen 

 was removed with forceps, using the dissecting microscope. The 

 pollen was scraped into a heap on the centre of the slip and 

 stained with methylated spirit in which a few granules of methyl 

 violet had been dissolved. This stain must not be too dark ; 

 it should be quite transparent, though violet in colour. This 

 process lasts about a minute, several drops being added at in- 

 tervals, and the slip is then placed on the hot plate. In the 

 next process watch carefully to see that the liquid is in every 

 case not completely evaporated. At the psychological moment 

 add a drop of unstained spirit ; repeat this, then add a drop 

 of turpentine ; repeat this three times, add a drop of balsam 

 and xylol, and cover with the thin glass. The cover-glass 

 should be placed on the edge of the slip, so as to be at the 

 same temperature when it is placed on the balsam, and it is 

 then less likely to hold air bubbles. The slip will be dirty with 

 stain and turpentine ; this can be removed when the slide is 

 cold with a rag dipped in spirit. 



N.B. — When the cover-glass is on, extinguish the lamp, and 

 let the slide cool with the hot plate. 



* Later on perfectly fresh pollen was experimented with, and found to 

 be equally good, so that it appears unnecessary to dry it. One of the 

 Compositae was used, stained with i'uchsin — a granule in methylated 

 spirit. 



Joura. Quekett Microscopical Club, Ser, 2, Vol. X., No. 65, November 1909. 



