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The biggest problem with sterile areas for tissue culture (plant or animal or bacteria) aren't usually with the setup itself. Your rubbermaid box with a HEPA filter was probably as good as any multi-thousand-dollar laboratory system.

The problems arise from two areas:

1) Air flow. You have to keep the outside air from blowing into your setup. The biggest causes of stray outside air are drafts (easy enough to detect and control) and people walking by the setup while it is in use. People walking by cause huge air currents--you wouldn't believe how much disturbance they unwittingly cause.

2) Sterile technique: there's a lot more to it than just wearing gloves. Everything in the hood has to be sterile, and that includes forearms and/or lab coats (arm gaiters are an easily sterilizable alternative). If UV lights are used to sterilize, they have to be the correct wavelength (about 285 nm--not 302 nm or higher). The lights also have to be less than 6 months old, and free of dust or other obscuring contaminants on the bulbs. The HEPA filter has to be clean--it also needs regular changing and servicing.

That's all in addition to not getting careless and failing to use proper sterile technique, not removing hands from the hood, not touching the work surface with arms or elbows, not sneezing or coughing into the hood, holding caps and lids down, removing wrappers from things correctly, and not moving too quickly within the hood (or in and out of it) because of the air currents generated.

It's really staggering the amount of work needed to keep even the most state-of-the-art sterile chamber or hood sterile during use.

(Been there done that for nine years and it was the part of my job I liked least)

I institute it exceptionally riveting.

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