Lab Final Review - Week #2 Flashcards
What is a stain (chemically)?
an organic compound made up of a benzene ring plus a chromophore and an auxochrome group
The ability of a stain to bind to macromolecular componenets like proteins or nucleic acids depends on?
The electrical charge found on the chromogen portion and the electrical charge of the organisms being stained
Acidic stains
Anionic; negative chromogen -> binds to positively charge components like proteins
Basic stains
Cationic; positive chromogen -> binds to negatively charged components like nucleic acids
What is a chromogen?
It is the colored compound in a stain
What time of stain is usually used for bacterial staining? Why?
Basic stains, because of the negative charge on bacterial surfaces
Simple staining
Use of a single stain, which produces a distinctive contrast between the organism and the background. It is useful in the visualization of the morphilogical shape an arrangement of an organism
Differential staining
Use of two contrasting stain; at least 4 chemical reagents are needed to accomplish this. It is useful in separating microorganisms into groups & visualization of structures
What are the 4 primary rules in preparing bacterial smears for staining?
- clean slides
- label slides
- smear bacteria
- heat fixation
How would you prepare a bacterial smear using a broth culture?
Use one or two loopfuls and spread to about the size of a dime
How would you prepare a bacterial smear using cultures from a solid medium?
dilute the culture by placing one or two loopfuls of water on the slide in which the cell swill be emulsified. Spread to the size of a nickel
Why is heat fixation of the prepared slide so important?
So that the bacterial smear does not wash away during the staining procedure. Heat fixating coagulates the bacteria and fixes them to the glass surface
How could a smear that is too thick yield a false result?
Because of the thickness it could prevent stain to be washed away that otherwise would be
What are the 3 most commonly used basic stains?
- methylene blue 2. crystal violet 3. carbol fuchsin
How are simple stains clinically relevant?
Although they are not differential or diagnostic, simple stains are a quick procedure that allow for determining whether the sample has the presence of a foreign bacterial pathogen