What is Tissue/Cell Culture?
Tissue/cell 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/cell 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 aseptic conditions.
Steps involved in Tissue/cell 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.
Major equipments involved in Tissue/cell culture for providing various conditions such as Aseptic conditions
Incubators
Incubation is a significant process for the growth of cell in the Tissue/Cell Culture. Growing cells require strictly controlled environment. Incubator provides the correct growth conditions, such as temperature, degree of humidity and CO2 levels in a controlled and stable manner. Incubators minimize contamination problems apart from offering energy efficiency and safe environment for cell lines. Some Incubators also have the facility to control oxygen level. Copper-coated incubators are reported to reduce the risk of microbial contamination within the Incubator due to the microbial inhibitory activity of copper. The inclusion of a bactericidal agent in the Incubator water trays will also reduce the risk of bacterial and fungal growth. However, there is no substitute for regular cleaning. CO2 Incubators provide best optimal conditions for Cell/Tissue Culture.
Shakers
Shakers are one of the important Tissue/Cell Culture equipments. Laboratory shakers including Incubated Shakers are used in a wide variety of scientific applications such as
Molecular biology,
Plant and animal cell culture,
Fermentation,
Genetic research,
Drug discovery and
Microbiology.
Centrifuge
Centrifuges are used routinely in Tissue/Cell Culture as part of the subculture routine for most cell lines and for the preparation of cells for Cryopreservation. Only low speed centrifuges are used generally at 20 degree C to avoid disruption of the separated bands of cells.The motor releases the heat which leads to the increase in temperature. Therefore, use of low temperature for centrifugation is recommended so that cells are not exposed to high temperature. The centrifuge should have a clear lid so that the condition of the load can be observed without opening the lid. This will reduce the risk of the operator being exposed to hazardous material if a centrifuge tube has broken during centrifugation. Care should always be taken not to over-fill the tubes and to balance them carefully. These simple steps will reduce the risk of aerosols being generated.
Laminar Air Flow
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
Seed Germinators
A seed germinator is Tissue/Cell Culture equipment. Using Seed Germinators, varying atmospheric conditions can be created within the chambers without the need of going outside. Typically, these create an environment in which light, humidity and temperature are controlled to provide optimum conditions for the germination of seeds. They are made of double walled structure having two chambers, one inner and one outer. The main components of a Seed Germinator include;
A germinating cylinder
A temperate controller
Seed Germinators are designed in such a way that they are able to control the crucial environmental parameters.
Microscopes
The Microscope is used for visualizing Tissue/Cell Cultures in situ. The cells in culture vessel remain at the bottom of the vessel and the medium floats above the growing cells. It is impossible to observe these cells under the ordinary Microscope; therefore, the Inverted Microscope is used for such purposes. The Inverted Microscope has the optical system at the bottom and the light source at the top, this arrangement helps to observe the cultured cells in the plates.