BUILDING A STERILE TRANSFER CHAMBER
Nan's Nook : Archives : Gloveboxes
In order to achieve a good success rate when making transfers, you must first construct a transfer chamber or “glove box.” This is simply an enclosed area that can be cleaned with disinfectants to keep out contaminants. Such a chamber can be designed in many ways and need not be elaborate to work well. Basically what you are striving for is an enclosure which provides a sterile, draft-free environment to work in while performing sterile culture work.
A good, inexpensive glove box can be constructed from a cardboard box, some aluminum foil and a sheet of plexiglass.
You will need the following materials:
Clean, sturdy cardboard box.
Razor knife or box cutter
Can of spray adhesive.
Roll of heavy duty aluminum foil.
Roll of duct tape.
A piece of plexiglass cut to approximately 10” x 14”.
Clean dish towel.
Obtain a good, clean, sturdy cardboard box with a removable lid. The kind paper
is shipped in is ideal and can be found at any printing shop or office supply
Use a small dish or bowl as a template and draw two circles about 5” in diameter on the front of the box. Cut out the two circular holes in the front side of the box (to allow you to insert your arms while making transfers). Also cut a square hole in the lid of the box slightly smaller than the sheet of plexiglass (approx. 9” x 13”). This hole will be covered with the sheet of plexiglass, effectively forming a “window” in the top of the box.
A razor knife or box cutter makes an ideal tool for cutting these holes in the box. Once you have the holes cut you can proceed to step II and prepare to apply the foil.
Wipe the inside of the box with alcohol and allow it to dry. After the alcohol dries liberally spray the entire inner surface of the box (sides, bottom and top) with the supplied Lysol aerosol. This kills any spores that are lying dormant on the surface of the cardboard. Once the Lysol has evaporated, spray a layer of adhesive in the bottom of the box and apply a sheet of aluminum foil to cover. Do not worry if the foil has a few wrinkles or creases. You just want to have a nonporous surface that is easy to clean and sterilize as well as being waterproof. Repeat this process on all sides of the inside of the box.
Fit the lid onto the box. Place the sheet of plexiglass over the hole in the lid
of the box and tape each side to the box with duct tape. Be sure to seal each
edge as you tape the plexiglass down to the box. This forms a nice, airtight
“window” in the top of the box which will allow you see into the cabinet with no
risk of contamination from airborne spores and bacteria.
Leave the lid of the box removable (do not tape) to allow easy access for placing your materials inside prior to performing sterile transfer work.
Take a small, clean dishtowel and drape it across the top of the box so that it
hangs down over the two hand holes in the front.
Run a strip of duct tape along the edge of the towel taping it to the lid of the box along the edge of the plexiglass.
The towel serves as a barrier to prevent drafts from entering the cabinet while you are working, effectively keeping contaminants out.
Do not tape the lid down. Leave the lid removable so it will be easier later when you need to load materials into and out of the sterile cabinet.
Note from Nan - I like a clean plastic sheet instead of cloth
Congratulations! You now have a
completed, fully functional sterile transfer cabinet that only cost a few
bucks.... Spray inside of chamber with Lysol before each use but....ALLOW
ALCOHOL FROM LYSOL to EVAPORATE COMPLETELY!!! before lighting any burners or
lighters inside of box... unless you like the "hairless" look and don't mind
your arms stinging like hell for a day or two!
Note: You can save on the Plexiglas (which must be cut) but substituting some clear plastic film sheet (like a $2 plastic drop cloth) for the window and duct taping the plastic into place securely with no sags. I have done a lot of sterile work under draped plastic hoods and inside cardboard boxes; be clean in and around them and you can do intermediate mycology with only a few contam glitches
Also See: My Glovebox (Plastic and Duct Tape Creations) & OT Teks