Negative Staining Method and its Mechanism

Negative Staining Method and its Mechanism


  • Negative staining procedure is also called as Relief staining.
  • Some bacteria have non-ionic charge or slime layer on their surface.
  • Because of which the stain doesn’t penetrate inside the cell and the bacterial cells are not easily stained.
  • In case of such bacterial cells negative method is used for observing the morphological characters of the cell.
  • In negative staining method actually, the background is stained and the bacterial cell remains colourless.
  •  This is the only staining method in which bacterial cells are not stained but they are made visible against a dark background.
  • Generally, the acidic stain such as 10 % Nigrosin, Eosin, India ink and Congored is used.


  1. A grease-free slide is taken and a smear is made on the slide by using Nichrome wire loop.
  2. The slide is allowed to air dry.
  3. Here there is no Heat fixation step.
  4. After air drying a drop of stain is added on an end of a slide and a thin film of stain is made by using another slide.
  5. These film is allowed to air dry and observed under oil immersion.

Read more