1.
Culture Methods
Culture methods involve growing microorganisms on suitable culture media under appropriate conditions so that their growth
characteristics can be studied.
A. Types of culture media
1. Liquid media (broth)
Microorganisms grow throughout the liquid medium. Example: Nutrient broth.
2. Solid media
Contain a solidifying agent such as agar and allow microorganisms to form colonies. Example: Nutrient agar.
3. Selective media
Allow the growth of particular microorganisms while inhibiting others.
4. Differential media
Help distinguish microorganisms based on visible biochemical reactions, such as changes in colony appearance or medium color.
B. Common culture techniques
1. Streak plate method
A sample is spread over the surface of solid agar in a pattern that progressively separates cells, allowing isolated colonies to
develop.
2. Spread plate method
A measured sample is distributed over the surface of an agar plate to obtain separated colonies.
3. Pour plate method
A sample is mixed with molten agar and allowed to solidify. Colonies can develop within or on the agar.
4. Slant culture
Microorganisms are grown on the surface of agar in a test tube containing a sloping solid medium. It is useful for maintaining
cultures and observing growth characteristics.
C. What we observe in culture?
After growth, we can study:
Colony morphology – size, shape, margin, elevation, texture and pigmentation.
Growth pattern – amount and distribution of growth.
Biochemical characteristics – reactions produced by the microorganism.
Growth requirements – temperature, pH, oxygen requirements and nutrients.
2. Microscopic Methods
Microscopic methods are used to observe microorganisms or their structures using a microscope.
Since most microorganisms are too small to be seen clearly with the naked eye, microscopy helps study their size, shape,
arrangement and cellular structures.
A. Types of microscopic methods
1. Bright-field microscopy
The most commonly used method in basic microbiology.
The specimen is viewed against a bright background.
Staining is often used to increase contrast.
2. Dark-field microscopy
The background appears dark while the microorganism appears bright.
Useful for observing very thin organisms that are difficult to see using ordinary bright-field microscopy.
3. Phase-contrast microscopy
•Converts differences in light passing through the specimen into differences in contrast.
•Useful for observing living, unstained cells.
4. Fluorescence microscopy
•Uses fluorescent dyes or labels.
•Structures containing the fluorescent label appear brightly against a darker background.
5. Electron microscopy
•Uses electrons instead of visible light and provides much greater resolution.
•TEM is mainly used to study internal ultrastructure.
•SEM is mainly used to study surface morphology.
Culture vs Microscopic Methods
Culture Methods Microscopic Methods
Microorganisms are grown on/in culture media Microorganisms are observed using a microscope
Used to obtain isolated colonies Used to study cell morphology and structures
Helps study growth and biochemical properties Helps study size, shape and arrangement
Can help obtain pure cultures Usually does not provide a pure culture by itself
Examples: bright-field, phase-contrast and fluorescence
Examples: streak, spread and pour plate
microscopy
Easy way to remember:
Culture method = “Grow the microorganism and study its
growth.”
Microscopic method = “See the microorganism and study its
morphology.”