SECONDARY STAINS- when gram stain doesn't work
1. Acid Fast Stain= Ziehl-Neelsen Technique
-Acid fast bacteria= pink RODS= Mycobacterium smegmatis
-NONacid-fast bacteria=blue COCCI =Staphylococcus epidermis
ie. Mixed culture, NOT pure culture
Acid Fast 
Acid Fast
Acid Fast 2. Endospore Stains
a. 2 Types of Staining Techniques
1. Crystal Violet Method= negative stain
a. Vegetative cells= purple
b. Endospores= clear
2. Schaeffer-Fulton Method
endospores= green
vegetative cells= red
b. We are looking at bacteria: Bacillus subtillis in all these Endospore stains
Endospore Stain-Crystal Violet
Endospore Stain-Crystal Violet
Endospore Stain-Crystal Violet 
Endospore Stain-Crystal Violet
Endospore Stain: Schaeffer-Fulton
Endospore Stain: Schaeffer Fulton 
Endospore stain: Schaffer Fulton 
Endospore stain: Schaffer Fulton 
Endospore stain: Schaffer Fulton 
Endospore stain: Schaffer Fulton 3. Flagella Stain= secondary stain used to visualize flagella in motile bacteria
(sg. = flagellum) (pl= flagella)
a. flagella may or may not be present on bacteria-can have one, a few, or many
b. Leifson’s Technique= a method to placing bacteria with flagella on slide in order to NOT break their fragile flagella (bc regular smearing and heat fixing would break off flagella)
c. Bacteria seen in Flagella Stain= Proteus vulgaris =rod shaped bacteria
-note some cells with have multiple flagella, while some will not bc they were broken off during preparation
Flagella stain
Flagella stain
Flagella stain
Flagella stain
Flagella stain4. Capsule Stain = negative stain= dark background w. highlighted cell
background= dark from India ink
bacterial cells= red from safranin
capsules= clear/white= no stain
d. Prepared slides contain bacteria: Klebsiella pneumoniae
-seen as red rods with clear capsules and dark background
Capsule Stain
Capsule Stain 
Capsule stain 
Capsule Stain 
Capsule Stain 
Thank you for sharing your article on how to remove different kinds of stains. This has helped me lot.
ReplyDeleteaudrey
www.triciajoy.com