Method of Identifying Microorganism Flashcards
Study organisms in their living or natural state
Demonstrates motility
Direct Method
Types of Direct Method
Wet mount method
Hanging Drop method
Parts of Hanging drop method
Depreession (well) slide
Cover slip
Petroleum Jelly (seal)
Drop of liquid with specimen
Examine organisms in their fixed or stained
state
Indirect Method
Types of Indirect Method
Smear preparation
Staining method
Specimen for smear preparation
Solid specimen
Liquid specimen
What to do with liquid specimen in smear preparation
air dry
heat fix
Attach or secure microorganisms on the glass slide
Kill the microorganism
Heat Fix
artificial coloring of the smear or microorganism
with dyes.
Staining Method
Make organism more visible
For identification and classification of
microorganisms
To emphasize special bacterial structure
Staining
2 types of staining
Simple staining
Differential staining
Utilizes only one dye or stain
Simple staining
Employs 2 dyes
Differential staining
Stains in differential staining
Gram Stain
Acid Fast Stain
Components of Gram Stain
Crystal Violet
Grams Iodine
Acid Alcohol
Safranin
Color of Gram Negative
Pink or Red
Color of Gram positive
Purple or Blue
All cocci are gram (+) except?
Neisseria
Branhamella
Veilonella
Moraxella
All Bacilli are gram (-) except?
Bacillus
Lactobacillus
Listeria
Actinomyces
Corynobacterium
Clostridium
Mycobacterium
Erysipelothrix
Nocardia spp
Theories of Gram Staining
Mg RNA
Kaplan-Kaplann
Stearn-Stearn
❖Gram (+) organism’s cell wall contains Mg RNA
❖ Gram (-) organisms cell wall do not contain Mg RNA
Mg RNA
❖ Gram (+) organisms are less permeable to decolorizer
❖ Gram (-) organisms easily decolorized by alcohol
Kaplan- Kaplan
❖ Gram (+) organisms have lower isoelectric point making them more
acidic
❖ Gram (-) organisms have higher isoelectric point making them more
basic
Stearn- Stearn
Gram stain cannot stain?
Mycobacteria
Mycoplasma
Rickettsiae
Chlamydia
Components of Acid Fast Stain
Carbol Fuchsin
Acid Alcohol
Methylene Blue
2 methods of Acid Fast stain
Zhiel-Neelsen Method
Kinyoun Method
Acid fast positive color
Red
Acid Fast negative
Blue
active detergent or surfactant in acid fast staining
Tergitol
artificially prepared material that provides the
nutritional and environmental requirements for
bacterial growth
Culture media
Basic Ingredients of culture media
Beef Extract
Peptones
Salts
Water
Agar
Types of Culture media state
Solid
Liquid
Semi-solid
With solidifying agent. It provides firm surface
Solid media
No solidifying agent. Remains in liquid form
Liquid media