Hello
Any laboratory, in which tissue culture techniques are performed, regardless of the
specific purpose, must contain a number of basic facilities. These usually include the
following:
•
A general washing area
•
A media preparation, sterilization, and storage area
•
An aseptic transfer area
•
Environmentally controlled incubators or culture rooms
•
An observation/data collection area.
The media preparation area should have ample storage space for the chemicals, culture
vessels and closures, and glassware required for media preparation and dispensing.
Bench space for hot plates/stirrers, pH meters, balances, water baths, and media-
dispensing equipment should be available. Other necessary equipment may include air
and vacuum sources, distilled and double-distilled water, Bunsen burners with a gas
source, refrigerators and freezers for storing stock solutions and chemicals, a microwave
or a convection oven, and an autoclave or domestic pressure cooker for sterilizing media,
glassware, and instruments.
Under very clean and dry conditions, tissue culture techniques can be successfullyperformed on an open laboratory bench. However, it is advisable that a laminar flow hood or sterile transfer room be utilized for making transfers. Within the transfer area
there should be a source of electricity, gas, compressed air, and vacuum.
All types of tissue cultures should be incubated under conditions of well-controlled
temperature, humidity, air circulation, and light quality and duration. These
environmental factors may influence the growth and differentiation process directly
during culture or indirectly by affecting their response in subsequent generations.
Protoplast cultures, low-density cell suspension cultures, and anther cultures are
particularly sensitive to environmental cultural condition.
Typically, the culture room for growth of plant tissue cultures should have a temperature
between 15° and 30° C, with a temperature fluctuation of less than ±0.5°C; however, a
wider range in temperature may be required for specific experiments.
Regards
Ashwini