Staining roots for SCN

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Place the root samples in separate 100 or 150 ml glass beakers. Prepare 600 ml of a water-bleach solution by adding 100 ml household bleach (5% sodium hypochlorite or NaClO) to 500 ml distilled water. Add about 50 ml of water-bleach solution to each beaker containing a root sample.

Incubate roots in the water-bleach solution for 4 minutes, stirring with a metal spatula periodically. Rinse roots for 45 seconds or so in running tap water, then soak the roots in tap water for 15 minutes.

Pour off the tap water, rinse with tap water one last time, then add approximately 40 ml of distilled water and approximately 7 or 8 drops of the acid fuchsin stain to each beaker. Place beakers containing roots and stain on a hot plate and heat until boiling fairly well. Then remove from the hot plate and allow to cool enough so that they are comfortable to touch.

After cooling, carefully rinse roots in running tap water for approximately 1 minute, then carefully drain off most of the excess water. Add 10 to 20 ml of acidified glycerine to each beaker. Acidified glycerine is prepared by adding 5 to 10 drops of dilute 1.0 M HCl to regular glycerine.

To prepare the acid fuchsin stain

Add 3.5 g acid fuchsin and 250 ml glacial acetic acid to 750 ml distilled water. Stir well.

Stained roots with nematodesSCN feeding on roots viewed under the microscope after staining