Schaeffer-Fulton stain

Schaeffer-Fulton stain

1. Pure culture of the suspect strain to be studied.

   • The culture is subjected to extreme temperatures for 24 hours to stimulate the microorganism to sporulate.
     • Place a few drops of sterile physiological solution on a clean slide.
       • Then a small portion of the crop is taken and a fine spread is carried out.
         • It is left to dry and it is fixed to the heat.
           • Primary stain: cover the slide with 5% aqueous solution of malachite green (A filter paper can be placed on the foil).
             • Heat over the Bunsen burner flame until steam is released and remove the flame. Repeat the operation for 6 to 10 minutes.
               • Remove the filter paper (if it was inserted) and wash with water.
                 • Counter stain:cover the slide with 0.5% aqueous safranin for 30 seconds.
                   • Wash with tap water; blot dry.
                     • Examine the slide under microscope for the presence of endospores. Endospores are bright green and vegetative cells are brownish red to pink.