Scientists study viruses by implanting them into cells derived from a tissue culture. Another use of tissue cultures is the formation of somatic cell hybridisation, in which two dissimilar cells are fused to produce a single hybrid cell, often used in genetic mapping. Tissue culture techniques are often used in plant science to investigate the effects of gene regulation and totipotency, the ability of plant cells to differentiate into different tissue types. Setting up a tissue culture lab requires proper contamination control and design.
- Skill level:
Things you need
- 150 square feet of space
- Autoclave or pressure cooker
- Distilled or deionised water
- pH meter
- Microwave oven (optional)
- Vacuum pump or aspirator
- Culture dishes
- Graduated cylinders
- Humidifier and dehumidifier
- Laminar flow hood or box with one open side
Write a general plan that details the location, size, lighting, electricity and general aseptic capabilities of the location for your tissue culture laboratory. You can set up a small lab in the basement of a house, a garage or a remodelled office. The minimum area required for proper media preparation, transfer and growth areas is 150 square feet.
Check with local authorities about zoning and building permits. Cover the walls and ceilings with a water-resistant material and ensure that the location has at least 100 amp electrical service.
Design the layout of your laboratory space considering the aseptic needs of each area. The cleanest areas should be those for culturing and growing tissue samples. This area and the aseptic transfer area should be located away from the main entrance. Locate the media prep area, glassware-washing area and storage area away from the cleanest areas.
Install an autoclave, a laboratory device that sterilises glassware using high levels of heat, or install and use a pressure cooker to sterilise glassware. Both hot and cold water should be available with access to distilled or deionised water.
Install the following in the media preparation area: refrigerator/freezer, balances that can measure to 2 mg, hotplate, pH meter and an aspirator or vacuum pump. You may include a microwave oven for heating culture media. Microscopes, beakers, flasks, culture dishes, graduated cylinders, forceps, pipettes, scalpels and spatulas are basic laboratory equipment you will need.
Assess the needs of the culture area in terms of temperature, humidity and lighting. You should generally keep the laboratory temperature between 24.4 degrees C +/- -16.7 degrees C. Control the relative humidity of the area with either a humidifier or dehumidifier; cultures can dry out if the room's relative humidity is less than 50%.
Maintain a separate area for aseptic transfer of cultures. You should install laminar-flow hoods in this area. If installing a hood is not possible, you can decrease contamination by transferring your tissue cultures under a box with only one open side.
Tips and warnings
- When propagating plants, you should ensure that your plant specimens are free of contamination. You should obtain your plant specimens from organisations such as the Department of Agriculture, agricultural universities or privately owned repositories that produce plants that have been tested for the presence of pathogens.
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