MICROBIOLOGY
SPOTTERS PART-II
-Dr. Babita Fageria
Incubator
• Used in laboratories for growth & maintenance
of micro-organisms & cultures
• It has wide range of applications including cell
culture, pharmaceutical studies,
haematological studies, biomedical studies.
• It can be used in stem cell research
Hot Air Oven
• Dry heat sterilization performed on dry material & on
substances that do not melt or catch fire under high
temperature.
• Principle: heating the materials at 160°C for 1 hour, allows
destruction of vegetative cells as well as spores
• Uses: to sterilize materials like glasswares, metal
equipments, latex powders etc.
• Biological indicator- Bacillus atrophaes
Inspissator
Principle: heating article on 3 successive days at 80-85 °C for 30 minutes each.
First exposure kills all the vegetative cells, and withdrawl of heat intermittently
stimulates the spores to germinate which can be killed on subsequent exposure.
So also called as intermittent sterilization.
Use: for sterilizing egg and serum based media, which can be disintegrated by
high heat of autoclave e.g. Lowenstein Jensen media, Loeffler’s serum slope
Laminar Flow
• This device creates a sterile
environment with the flow of air
through a High Efficiency
Particulate Air (HEPA) filter &
short wave ultraviolet germicidal
lamp that sterilises the work station.
• Use:
Clinical microbiology-used to carry
out laboratory procedures that are
sensitive to contamination, such as
processing of biological samples for
bacteriological culture.
Others- also for experiments related
to plant tissue culture & genetic
transformation
Water bath
Principle: Sensor in the device transfers water temperature to a reference
value which is then amplified. Control system generates a signal for the
heating system which heats the water to the desired temperature.
Use: Primarily used for heating samples under a controlled temperature.
Also suitable for heating chemicals that might be flammable under direct
ignition.
Centrifuge
Centrifuge
• A device that allows the rotation of an object about a single axis where
an outward force is applied perpendicularly to the axis.
• Laboratory centrifuge is motor based and allows the rotation of liquid
samples resulting in the separation of the components of the mixture
viz. sediment and supernatant.
• Principle: Works based on density gradient where denser particles
settle at the bottom while lighter ones remain suspended at the top.
• Uses:
Separation of cell organelles
Separation of blood components in immunology & hematology
Stool concentration techniques
Nucleic acid extraction for PCR
Separation of isotopes
Bunsen burner/ Spirit lamp
It is a standard tool used in clinical laboratories named after Robert Bunsen.
It is a gas fueled single open flame.
Principle: Once the burner is connected to gas supply, the gas is forced by its
own pressure so that it reaches the top where the flame is ignited using a lighter
or similar tool.
Uses: Commonly used for processes like sterilization and heating liquids etc.
In clinical microbiology laboratory, it is mainly used for sterilizing
inoculating loop, wire, forceps and mouth of test tubes and flasks.
INDOLE TEST
• PRINCIPLE:
• This test demonstrates the ability of
certain bacteria to convert the amino
acid tryptophan to indole, which
accumulates in the medium.
• Tryptophan is decomposed by an
enzyme tryptophanase produced by
certain bacteria.
Tryptophanase
• Tryptophan
Indole
• Interpretation:
• Indole positive: Red coloured ring
• e.g. E. coli, Shigella, Edwardsiella,
Proteus sp. Other than P. mirabilis
• Indole negative: Yellow coloured ring
• e.g. Klebsiella sp, Enterobacter sp.,
Serratia, Hafnia sp., Proteus mirabilis
METHYL RED TEST (MR)
PRINCIPLE
The methyl red test is employed to
detect the productiion of sufficient
acid during the fermentation of
glucose so that Ph of the medium
falls and it is maintained below 4.5
INTERPRETATION
MR POSITIVE – bright red
colour
eg: [Link], Shigella sp, Edwarsiella
sp, yersinia sp, Listeria
monocytogenes.
MR NEGATIVE: yellow colour
eg: Klebsiella, Enterobacter sp,
Serratia, Hafnia sp.
CITRATE TEST
PRINCIPLE:
This is a test for the ability of an
organism to utilize citrate as the sole
source of carbon for growth and
ammonium salts as the sole source
of nitrogen with resulting alkalinity.
INTERPRETATION:
CITRATE POSITIVE: Blue colour
e.g. Klebsiella sp., Enterobacter sp.,
Serratia sp., Hafnia sp., Salmonella
sp. except S. Typhi, Citrobacter sp.
CITRATE NEGATIVE: Green
colour
e.g. E. coli, Edwardsiella sp.,
Shigella sp., Salmonella Typhi
