Glove boxes, flow hoods and the economical glove bag are important tools for creating a sterile environment.
This is an alternative to making a glove box and is several times more effective when using ozone a sterile gas to inflate the bag.
We used a 3D printer to make the rings that hold the gloves to the bag but, anything that is semi-round that the cuff of the gloves can fit around and a rubber band will cling to work fine In the first iteration we used the metal bands from mason jars. Those might be too small for some peoples hands and if there's no 3D printer for miles where you live a 2" section of corrugated pipe should work as the rings. Theory:
Creating a pure culture of an organism means you don't want any extra contaminants like mold, bacteria or any living thing (besides what you want to grow) that invisibly swims and crawls all around us. This proves to be as tricky as creating a 100% dust free environment and even more so since tiny organisms are very hard to neutralize and stick to any surface.
Alternative methods in a tissue lab include: Filtering air, aerosol of liquid chemicals, inert gases and chemically reactive gases.
We choose a method for sterilization that scales down to fit inside an envelope, uses a reactive gas but, is safe to the user and is also convertible to a liquid propellant sterilization, filtered air or inert gas method.
Since we're inflating a bag there is always positive pressure inside due to slow deflation. This means tiny cracks and holes aren't as troublesome since nothing can float in.
Using a commercially available Ozone pump is an affordable method for producing a sterile gas. It only takes tiny concentrations to do it's job and is also an irritant to humans. So when using ozone be sure you're in a well ventilated space or outside. Ozone also quickly breaks down back into oxygen so, after enough time has elapsed (24 hours) what will be left is sterile lifeless oxygen that sensitive organisms like fungal spores can be introduced and unaffected.
Ozone is a harmful gas. It should always be used in a ventilated area or outdoors. Ozone destroys organic compounds and can deteriorate items such as rubber. Please read the Material Safety Data Sheet on Ozone (O3)
- Large clear plastic bag
- elbow length gloves (rubber gloves will crack and fall apart if you plan to use ozone) search for poly elbow gloves for ozone resistance
- A plastic ring large enough for your hands to fit through
- Rubber bands that will hold to your rings
- A clip or tie to close the bag Print files are located here:
http://www.thingiverse.com/thing:114023 A decent ozone pump on amazon
There's even cheaper ones on eBay for about $30
And if you need help printing the rings, I offer 3D printing services here:
http://www.makexyz.com/printer/operator Assembly Steps:
1) Put the opening to a glove (poly gloves work best) through the middle of the ring and wrap the cuff over the edge of the ring.
2) Push the ring and glove opening against the bag from the inside of a clear (poly) bag or clear trash bag.
3) Use a rubber band to secure the glove ring to the bag from the outside and cut an opening into the glove ring.