Neisser Stain
The Neisser stain is a differential stain in much the same manner that the Gram stain uses more than 1 stain. The bacteria will either stain Neisser positive (purple) or Neisser negative (yellow).
Neisser Stain Procedure
Preparation:
Solution 1:
Part A: 0.1 g Methylene Blue, 5 mL 95% Ethanol, 100 mL Distilled Water.
Part B: Crystal Violet (10% w/v in 95% Ethanol) 3.3 mL, 6.7 mL 95% Ethanol, 100 mL Distilled Water
Mix 2 parts of volume of Part A and 1 part volume Part B: Prepare fresh monthly
Solution 2:
Bismark Brown (1% w/v aqueous) 33.3 mL
Distilled Water 66.7 mL
Staining Procedure
Prepare thin smears on the microscope slides and thoroughly air dry. Do not heat fix.
Stain 30 seconds with Solution 1;, rinse 10 seconds with water
Stain 1 minute with Solution 2; rinse well with water; blot dry
Examine under oil immersion at 1000x magnification with direct illumination (Brightfield) not phase contrast. Blue-violet is positive (either entire cell or intracellular granules; yellow-brown is negative
Video of Neisser Staining Procedure