Plant Tissue Culture Laboratory/Tissue Culture Room Planning and Design
Plant Tissue Culture Laboratory/Tissue Culture Room Planning and Design
September 22, 2023
The ideal tissue culture laboratory should be located in a quiet, clean location away from the source of pollution, preferably upwind of the prevailing wind direction throughout the year to minimize pollution. A mass-produced tissue culture lab is best located in a convenient location where the culture products can be transported easily.
Basic principles of laboratory setup PART 01
The construction of laboratories should consider two aspects of the problem: one is the nature of the experiment, that is, whether it is productive or research, is basic level or higher level; The second is the scale of the laboratory, the scale mainly depends on the funding and experimental nature.
Regardless of the nature and size of the laboratory, the basic principles of laboratory setup are: scientific, efficient, economical and practical. A tissue culture laboratory must meet three basic needs: laboratory preparation (culture-medium preparation, dishwashing, sterilization of culture-medium and culture-vessel), aseptic operation, and controlled culture.
Basic laboratory PART 02
The basic laboratory includes preparation room, washing and sterilization room, aseptic operation room, culture room, buffer room, which is the basic condition necessary for tissue culture experiment.
Preparation room (Chemistry laboratory)
Carry out all the preparatory work related to the experiment: complete the storage, weighing, dissolving, utensil washing, preparation and subassembly of the medium, sterilization of the medium and the culture utensil, pretreatment of the culture material, etc.
Requirement:
The best design is around 20 square meters. Spacious and bright, in order to place multiple experimental platforms and related equipment, convenient for many people to work at the same time; At the same time, good ventilation conditions are required to facilitate gas exchange; The laboratory floor should be easy to clean and non-slip treatment should be carried out.
Washing and sterilization room
Complete all kinds of equipment washing, drying, preservation, sterilization of medium, etc.
Requirement: Washing sterilization room according to the size of the workload to determine its size, the general area is controlled in 30-50m2. Set up a special sink on one side of the lab for cleaning glassware. The central test bench should also be equipped with 2 sinks for cleaning small glassware. Invest in a bottle washing machine if the workload is heavy. Prepare 1 or 2 wash tanks specially for washing glassware with high cleanliness requirements. The floor should be resistant to moisture and well-drained.
Equipment: Pool, drop rack, central experimental platform, autoclave, ultrasonic cleaner, drying sterilizer (such as oven), etc.
Sterile operation room (inoculation room)
It is mainly used for disinfection of plant materials, inoculation, transfer of culture, subgeneration of test-tube seedlings, preparation of protoplasts and all technical procedures requiring aseptic operation. It is the most critical step in the research or production of plant culture in vitro.
Requirement:Inoculation room should be small rather than large, generally 7 ~ 8 square meters (10-20 square meters), its size according to the needs of the experiment and environmental control of the degree of difficulty. It requires good sealing, dry and quiet, clean and bright, and can maintain sterility for a long time, so it is not easy to be located in a place prone to moisture; The floor, ceiling and walls should be sealed and smooth, easy to clean and disinfect; Configuration pull the door to reduce the air disturbance when opening and closing; In order to facilitate disinfection, the ground and inner walls should be made of waterproof and corrosion resistant materials; In order to keep clean, the aseptic room should prevent air convection. The inoculation room requires that 1 ~ 2 ultraviolet sterilizing lamps should be hoisted in the appropriate position for irradiation sterilization. It is best to install a small air conditioning, so that the room temperature is controllable, so that the doors and Windows can be closed, reduce the convection with the outside air.
Equipment: Ultraviolet lamp, air conditioning, anatomical mirror, sterilizer, alcohol lamp, inoculation equipment (inoculation tweezers, scissors, scalpels, inoculation needle), experimental bench, and shelves, as well as a clean table and a complete set of sterilization inoculation equipment, drugs, etc.
Culture room
In vitro plant materials inoculated into culture bottles and other vessels are cultured under controlled conditions, whether in research laboratories or places where productive experimental materials are cultured.
Requirement: About 10-20square meters, the size of the culture room can be based on the need for the size of the incubator, the number, and other ancillary equipment, its design to make full use of space and save energy for the principle. The basic requirements are to be able to control light and temperature and to maintain a relatively sterile environment. Therefore, the culture room should be kept clean and moderately dry; In order to meet the demand of gas for the growth of plant culture materials, ventilation devices such as exhaust Windows and ventilation fans should be installed in the culture room. In order to save energy and space, suitable culture stands should be equipped and fluorescent lighting should be installed.
Equipment: Culture frame (temperature control, light control and humidity control), shaker, rotary bed, automatic time controller, UV lamp, light incubator or artificial climate box, biochemical incubator, side table experiment platform for filming the growth of culture, dehumidifier, microscope, hygrograph, air conditioning, etc.
Aseptic operation equipment PART 03
Inoculation box
Is the earliest use of the simplest aseptic device, the main body is a glass box cover, the entrance has a sleeve cover, equipped with ultraviolet lamp and fluorescent lamp, the use of aseptic room requirements.
Cleaning bench
The operating table is a semi-open area, with the advantages of convenient and comfortable operation. Through continuous blowing of filtered air, microorganisms larger than 0.03um in diameter are difficult to stay in the operating space of the table, maintaining a better aseptic environment.
Culture equipmentPART 04
Culture equipment refers to the facilities and equipment used for the culture of plant cells, tissues and organs with different specifications and control precision selected according to the needs.
Culture rack
The current facility for plant propagation and culture in all plant tissue culture laboratories. Low cost, flexible design, can make full use of training space, easy to operate, maximize the use of training space as the principle. Generally there are 4-5 layers, the interval between the layers is 40-50cm, the light intensity can be determined according to the characteristics of the cultivated plants, generally each rack is equipped with 2-4 fluorescent lamps.
Incubator
For cell culture, protoplast culture and other experiments requiring precise culture, light incubator, CO2 incubator and humidity control incubator can be used for culture.
The main difference between cell culture techniques and other general laboratory work is the requirement to maintain sterile operation and avoid the influence of microorganisms and other harmful factors. At present, the widespread use of the ultra-clean table has greatly facilitated the work of tissue cell culture and made it possible for some conventional laboratories to be used for cell culture.