Tech. D:6 



fuchsin method (Goodey 1937) was developed (McBeth, Taylor, and Smith 

 19hl) in order to eliminate the necessity for dehydrating the tissue. 



Goodey' s formula for staining is used as follows: 



Phenol, pure crystals 20 g. 



Lactic acid 20 g. 



Glycerine hO g. 



Distilled water 20 ml. 



Acid fuchsin or cotton blue ^ ml. (1 g. to 100 ml. water) 



The washed roots are boiled in the staining solution for one minute. 

 Wash in tap water to remove excess stain and place in lactophenol solu- 

 tion vmtil cleared. The same lactophenol formula is used for clearing 

 that is used for staining, except that no stain is added. Clearing 

 requires from one to several days, depending upon the type of roots and 

 intensity of the stain. After clearing, the roots can be left in lac- 

 tophenol or in glycerine for examination. If permanent mounts are 

 desired, the material can be rtounted directly in glycerine. 



In some cases, better results are obtained by using a weaker stain con- 

 centration, -g- ml. of stain solution in 100 ml. of clear lactophenol. 

 Boil the plant parts for two minutes in clear lactophenol, then for one 

 minute in the stain, or for two minutes in the stain. 



Leaves stained in cotton blue lactophenol (Franklin 19U9) 



Prepare a 0.1 per cent solution of cotton blue in lactophenol (stock 

 lactophenol for this purpose consists of phenol, 20 g.; lactic acid, 

 20 g.; glycerine, UO g.; and water, 20 g.). 



Plunge leaves into boiling 0.1 per cent cotton blue lactophenol and 

 keep submerged in the gently boiling stain for 3-5 minutes. Leave in 

 the stain, which is permitted to cool. Remove tissues and wash in run- 

 ning water or in $0 per cent alcohol to eliminate excess stain. Clear 

 in lactophenol or in concentrated phenol. Concentrated phenol acts 

 more quickly and removes the chlorophyll. More or less permanent 

 mounts can be made by transferring the plant material through 50 and 70 

 per cent alcohol to iso-butyl alcohol and moimting in euparal. If 

 euparal gets cloudy before mounting is completed, warm the slide gently 

 until cloudiness disappears. The American equivalent for euparal is 

 diaphane. Mounting in this medium may be from either 9^ per cent or 

 absolute alcohol, but one should work quickly to avoid clouding. 



Itoots of some plants, particularly of trees, are not satisfactorily 

 stained by the preceeding methods because of oils and other materials 

 present in the roots. These types of roots will reouire staining and 

 sectioning by standard methods. The procedures recommended for demon- 

 strating f\ingi and bacteria within plant tissues are also useful to 

 demonstrate nematodes. These methods are to be found i/i other refer- 

 ences (e.g. fiawlins 1936). 



