1) Crystal Violet  
2) Iodine
3) Alcohol
4) Safranin 

Gram Stain Procedure

Gram stain is a multi-step staining procedure that differentiates bacteria by the structure of their cell wall

Differential staining utilizes multiple different dyes to allow us to distinguish between different types of organisms. 

Differential Stains

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Gram Stain

Acid-Fast stains are used to determine whether or not a sample contains a *Mycobacterium* species, which have mycolic acid in their cell walls. Cells that contain the waxy mycolic acid will remain red like the primary stain, carbolfuchsin.

Acid-Fast Stains

Capsule staining often utilizes negative staining techniques like India Ink or Nigrosin to color the background, in order to visualize the encapsulated cells. Positively charged counterstains can also be used to view the cell with a clear capsule.

Capsule Stains

The Schaeffer-Fulton method is the most common method for viewing spores in a specimen. This method uses heat with the primary stain, malachite green, washed with water, and then counterstained using safranin. If endospores are present, green will remain visible in the cells. 

Endospore Stains

In flagella staining, a mordant, like tannic acid, is used to thicken the flagellum, if present, followed by pararosaniline or basic fuchsin to stain them.

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