1.4.  Greenhouse

The greenhouse facility is required to grow parent pants and to acclimatize in vitro raised plantlets. The size and facility inside the green house vary with the requirement and depends on the funds available with the laboratory. However, minimum facilities for maintaining humidity by fogging, misting or a fan and pad system, reduced light, cooling system for summers and heating system for winters must be provided. It would be desirable to have a potting room adjacent to this facility.

 

1.5. Equipments and apparatus

1.4.1. Media preparation area

•  benches at a height suitable to work while standing

•  pH meter is used to determine the pH of various media used for tissue culture. pH indicator paper can also be used for the purpose but it is less accurate. The standard media pH is maintained at 5.8.

•  hot-plate-cum-magnetic stirrer for dissolving chemicals and during media preparation

•  an autoclave or domestic pressure cooker is crucial instrument for a tissue culture laboratory. High pressure heat is needed to sterilize media, water, labware, forceps, needles etc. Certain spores from fungi and bacteria can only be killed at a temperature of 121°C and 15 pounds per square inch (psi) for 15-20 min. A caution should be taken while opening the door of autoclave and it should be open when the pressure drops to zero. Opening the door immediately can lead to a rapid change in the temperature, resulting in breakage of glassware and steam burning of operator.

•  plastic carboys for storing distilled water required for media preparation and final washing of labware.

•  balances near dry corner of the media room. High quality microbalance are required to weigh smallest of the quantities. Additionally a top pan balance is required for less sensitive quantities.

•  hot-air oven to keep autoclaved medium warm before pouring into vessels. It is also used for the dry heat sterilization of clean glassware like, Petridishes, culture tubes, pipettes etc. Typical sterilizing conditions are 160-170 °C/1hr.

•  Dishwasher for cleaning glass pipettes in running water

 

Figure 2.4: pH Meter and Magnetic stirrer-cum-heater

    

Figure 2.5: Autoclave with accessories and High Precision Weighing Balance

1.4.2. Storage area

•  a deep freezer (-20°C to -80°C) / refrigerator for storage of enzyme solutions, stock solutions plant materials and all temperature-sensitive chemicals.

•  microwave oven to melt agar solidified media

•  Upright and inverted light microscope with camera attachment for recording the morphogenic responses from various explants, calli, cells and protoplasts. Inverted microscope gives the clear views of cultures settled at the bottom of Petridishes.