Autoclave:

It is an instrument used to carry out sterilization of nutrient media, tissue culture glassware,
medical supplies, biological wastes etc.

The autoclave was invented by Charles Chamberland in 1884.

Principle:

When water is heated in a closed container, steam is produced under pressure. According to
Boyle’s Law, when volume is kept constant then temperature is directly proportional to
pressure. In this way, incoming steam increases pressure inside autoclave leading to
simultaneous increase in temperature. At 15 Psi (Pounds per square inch) a temperature of
121⁰C is achieved. When this temperature is maintained for 20 minutes, it coagulates cell

Autoclave uses a combination of high temperature, high pressure and time in order to kill
microorganisms and spores.

To be effective, the autoclave must reach and maintain a temperature of 121° C for at least 20
minutes by using saturated steam under at least 15 psi of pressure.

Tape indicators are used to validate autoclave process. These tapes have heat sensitive,
chemical indicator markings which changes color when exposed to temperatures of 121°C.

Tape indicators are typically placed on the exterior of the waste load. If the temperature
sensitive tape does not indicate that a temperature of at least 121°C was reached during the
sterilization process, the load is not considered decontaminated.

Tape indicators are not designed nor intended to prove that organisms have actually been
killed. They indicate that a temperature of 121°C has been achieved within the autoclave.

Dry sterilization:

Dry steam of higher temperature (containing less than 3% moisture) is used.

It requires more time for sterilization.

Exposure to the higher temperature may be harmful to the instrument.

Wet sterilization:

Moist steam of higher temperature (containing more than 3% moisture) is used.

It requires less time for sterilization.

Applications:

Used to decontaminate certain biological waste.

Sterilization of nutrient media, tissue culture glassware and medical supplies.

 Autoclave

Autoclave (Interior)

Laminar Air Flow

It is also known as ‘tissue culture hood’.

It is simply a workbench designed to prevent contamination.

Principle: In a laminar flow hood the air is passed through a HEPA (High Efficiency
Particulates Air) filter which has a pore size of 0.3 micro meter and hence removes all
airborne contamination like bacteria and spores to maintain sterile conditions.

The cabinet is usually made of stainless steel with no gaps or joints where spores might
collect.

In a laminar flow hood the air is passed through a HEPA (High Efficiency Particulates Air)
filter which has a pore size of 0.3 micron and hence removes all airborne contamination like
bacteria and spores to maintain sterile conditions.

Laminar hoods come in two configurations: Horizontal flow hood and Vertical flow hood.

Laminar flow cabinets have a UV-C germicidal lamp to sterilize the interior and contents
before usage to prevent contamination of experiment.

Germicidal lamps are usually kept on for 15 minutes to sterilize the interior and no contact is
to be made with a laminar flow hood during this time.

It is important to switch UV light off during use, to limit exposure to skin and eyes as
ultraviolet light emissions can cause cancer and cataract.

Glass sheid
HEPA filter

Laminar air flow cabinet