Plants in tissue culture are very vulnerable to disease. Everything has
sterile; the tissue, the containers, the tools, and the medium. Here’s
|Figure 10 Scientists and housewives use similar
Using a Pressure Cooker as an Autoclave
- Check the pressure cooker seal. If stiff or damaged, do not use the pressure
- Place items on a rack inside the pressure cooker.
- Add water to a level below the rack.
- Grease the lugs with petroleum jelly.
- Close the pressure cooker, making sure that the seal is well seated.
- Place the pressure cooker on high heat.
- Begin timing when a steady stream of steam emerges from the vent.
- After 15 minutes, turn the heat off and let cool before opening the pressure
follow the manufacturer’s directions for “canning” fruit and other
- Opening a pressure cooker while still warm is very dangerous.
- High pressure and hot steam can kill or maim.
Sterilizing Instruments and Other Items
Sterilizing Your Homemade Glove Box
tweezers, forceps, and scalpels in a wide-mouthed jar, business end
down, and cover with a doubled piece of aluminum foil. (These items can
resterilized inside the glove box by washing in bleach solution and
rinsing with sterile water.)
- Wrap single-edged razor blades (if used) in aluminum foil and autoclave
in the pressure cooker for 15 minutes.
jars covered with a doubled piece of aluminum foil. When the
pressure cooker is cool enough to open, snap a rubber band around the
- Sterilize water in glass jars with lids on loosely. Screw the lids down as
soon as the pressure cooker is opened.
paper inside a paper bag and autoclave in the pressure cooker above
water level. After sterilization, dry the bag in an oven set at 80°C.
unwrap the papers until needed.
These directions will work for both the inexpensive cardboard-plastic-film glove
box and the more expensive plastic glove box.
- Tie back your hair, roll up your sleeves, and remove your watch and other
- Collect and organize all the items you will need inside the cabinet.
- Close and seal the homemade glove box. (Plug in the HEPA filter fan, if
The active cabinet will not be tightly sealed like the passive one. (Air needs to
Note: The active cabinet will not be tightly sealed like the passive one. (Air needs to
- Wash your hands thoroughly with the Pert (or other disinfectant).
- Put on a pair of surgical gloves.
- Insert your gloved hands through the sleeves.
- Sterilize the inside of the cabinet, including your gloved hands, by spraying
with 70% alcohol and wiping dry with sterile tissue.
- Sterilize all jars and implements by spraying with 70% alcohol and wiping
dry with sterile tissue.
- Discard the used tissues in a small bag or box inside the glove box.
- If using the active glove box, do the following:
Note: From this point on, do not remove your gloved hands from the glove box.
If you must, close all jars first and resterilize everything before starting to work on
- Sterilize the inside of the inner cabinet, including your gloved hands,
by spraying with 70% alcohol and wiping dry with sterile tissue.
- Turn on the HEPA filter fan.
- Pass the items through the inner chamber.
Sterilizing Plant Materials
- Inside the homemade glove box, sterilize all plant material in diluted
domestic bleach solution containing 1 drop of detergent as a surfactant.
- Put plant pieces in a jar containing the bleach for 10–20 minutes. Agitate
- Transfer the tissue in the bleach solution to the sterile area.
- Discard the chlorine solution.
- Rinse plant pieces twice with sterile water.