To prepare suspension, pick small amount of growth from 18-24 h plate (equivalent to 1 mm colony). Do not pick up agar. Suspend gently in 3 ml distilled water. (Flagella can be knocked off.) Suspension should be faintly opalescent.
To prepare slide, pass cleaned slide through blue part of burner flame several times to remove residual dirt. Cool slide, flamed side up, on paper towel. Mark wax line across slide to give area 2.5 x 4.5 cm. Place large loopful of suspension in center of slide adjacent to wax line. Tilt slide, letting drop run down center of slide to end. If drop does not run evenly, slide is dirty. Discard it. Air-dry slide on level surface.
R15. Coating Solution for V. vulnificus EIA Phosphate-buffered saline 100 ml Triton X-100 (a polyoxyethylene ether)20 µl
Mix Triton X-100 with PBS, pH 7.4.
R16. Crystal Violet Stain (for Bacteria)
1. Crystal violet in dilute alcohol Crystal violet (90% dye content) 2 g Ethanol (95%) 20 ml Distilled water 80 ml
2. Ammonium oxalate crystal violet.