Tissue culture is a biological method of asexual propagation where a very small piece of tissue (shoot apex, leaf section, or even an individual cell) is excised (which is known as Explants) and placed in sterile /aseptic culture in a test tube/petri dish/tissue culture container containing a special culture media.
A tissue culture is used for rapid multiplication of an organism. A very specialized laboratory with sterile/aseptic conditions is required for tissue culture. All the procedure involved in tissue culture should be carried out in a special ventilated cabinet and these conditions are provided by Laminar Air Flow.
The culture medium is a proper mixture of the nutrients (organic and inorganic), vitamins, sugars, minerals and plant hormones especially Cytokinin and Auxin for shoot and root initiation.
Steps involved in Tissue Culture are
- Explants ; excision of the plant parts.
- Place the explants in a tissue culture container/test tubes/petri dishes.
- Multiplication ; Tissue grows and produce small plants.
- Rapid multiplication by transfer of fresh culture.
- Transplanting the plant into some kind of acclimation container.
Laminar Air Flow provides a work area with aseptic/sterile conditions for the tissue culture. Laminar Air Flow has continuous displacement of air (it provides streamline flow of air) that passes through HEPA (High Efficiency Particulate Air) filter that removes the particulates from the air.
Laminar Air Flow are equipped with a UV lamp that should be turned on about 10-20 minutes before being used to sterilize the shell or cabinet or the surface of the Laminar Air Flow to avoid any kind of contaminations. Wipe down the surface with ethanol before and after each use.
Laminar Air Flow can be vertical and horizontal. In the Vertical Laminar Air Flow the air blows down from the top of the cabinet. Vertical Laminar Air Flow is best for working with hazardous specimens since the aerosols that are generated in the cabinet are filtered out before they are released into the surrounding environments.
Horizontal Laminar Air Flow are designed such that the air flows directly at the operator hence they are not useful for working with hazardous specimens but are best protection for Tissue/cell culture.
Laminar Air Flow provides aseptic/sterile conditions for the preparation of Culture Media. All the culture vessels, test tubes, pipette, tip boxes, stocks of sterile eppendorfs should be opened only in the Laminar Air Flow to avoid any kind of contaminations. Culture Media cannot be prepared outside the Laminar Air Flow because it can get contaminated and there would not be any result.
