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OfflineNo_Life_G33k
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Sanitation for the OCD
    #5865965 - 07/16/06 03:49 PM (17 years, 7 months ago)

Sanitation Q

I'm curious about how 'real' labs maintain sterility and the associated procedures.

1)How do you move items from an autoclave(or PC) to a hood or glovebox without contaminating them? I'm assuming that unless you're in a clean room, there are still mold spores and bacteria present in the air.

2)When you put items into a glovebox- how do you sterilize them- jars, markers, alcohol swab pacakages etc. I see the antechambers on the big commercial glove boxes, but no one seems to talk about sterility- all the info seems to revolve around gasses and vacuum pressure.

I usually douse everything in a cloud of 70% isopropyl until it's dripping wet, then heavily mist/spray inside the glovebox, then lift the lid a little bit and slip the item into the box.
This method seems to work well, except that it takes a long time for the alcohol fumes to evaporate, and it messes up lighters/packages/spores.
It also gets messy when you spray 6 jars still wet from the PC.
Anyway to refine my technique???

I have a small HEPA filter with a fan behind it to help evaporate the alcohol and also to provide slightly positive pressure.

(As a slight aside- I think that your sterile technique is almost as important as killing every last floating mold spore.)

3) Alcohol and paper towels. A lot of people seem to use alcohol and paper towels to wipe down work areas and tools. So my question(s) are
(a): if the entire towels is NOT soaked in alcohol- wouldn't it be a vector for contams??? The corner of the paper towel that didn't get alcohol was exposed to 'dirty' air and is then introduced into a clean glovebox. So do you have to soak the entire towel to be safe?
(b): If you have a paper towel that is soaked in alcohol and is inside a glovebox, and you are going to use it to wipe down the inside of a glovebox(or other work area/tools), do you need to turn the towel to a 'fresh' side when you wipe different walls?? In other words -does the alcohol lose effectiveness as you use it. Do the contaminants on area 'A' get transferred to area 'B', or do they die on contact with the alcohol?
(C) restated- does alcohol kill all the nasties it encounters and still retain that ability as long as the towel is wet with alcohol?? Could you use the same alcohol wipe for all the needles???

3) With regard to paper towels- do I need a new roll for each time I do sterile work? or can I put an opened roll in a rubbermaid to keep it 'clean' for next time??

4) I use a lot of bleach to sanitize things... how much of that bleach could be replaced with Hydrogen Peroxide??? can I completely eliminate bleach and just use h2O2? Will H2O2 kill everything that bleach will?


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Make Your Own Magnetic Stirrer! How-To Tek w/ pix
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InvisibleFooMan
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Re: Sanitation for the OCD [Re: No_Life_G33k]
    #5865986 - 07/16/06 03:54 PM (17 years, 7 months ago)

Anything else?  :smirk:

First off, your sterilization techniques are overkill IMO. Just clean the glovebox well with regular soap and water, then close it until you place your jars inside. Depending on what you are trying to accomplish, many things can be done in an open-air environment.


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Invisiblecreamcorn
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Re: Sanitation for the OCD [Re: FooMan]
    #5866068 - 07/16/06 04:28 PM (17 years, 7 months ago)

1) when you move items from the autoclave/PC, they're usually sealed containers of sorts (jars, syringes, etc) whose insides are sterile.  it doesn't matter if the outsides of them get dirty, keep critical parts (i.e., needles, innoc holes, etc) covered up until you're innoculating, and work quickly.  alcohol swab the needle, and the injection site.

2) not everything that goes into the govebox is sterile.  the glovebox itself doesn't need to be.  it works because the air inside is still, there's no drafts to get contams airborne where they can then land on things and cause problems.

3) (a) not a concern, (b) no, just wipe it doesn't need to be a fresh section of paper towel, the alcohol kills things, so even if they stuck onto the paper towel and you wipe them onto something else, they're dead already (c) yes, if its wet, its good.

4) bleach is a little more effective than peroxide.  you can use peroxide though.  iodine works well too.

and agreed the things you speak of are pretty overkill.  working on a clean surface with washed hands in a draft free room means a 90%+ success rate.  all that extra stuff is for the last few percent... and you'll never hit 100%, contams will still happen on occasion even with the most anal procedures.  so save yourself the trouble and effort and supplies. :smile:


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Invisiblefastfred
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Re: Sanitation for the OCD [Re: creamcorn]
    #5866536 - 07/16/06 05:58 PM (17 years, 7 months ago)

> you'll never hit 100%, contams will still happen on occasion even with the most anal procedures.

Very true!

In the lab 1:10 bleach is used for surface sterilization and 70-80% isopropyl alcohol is used for sterilizing tools. Gloveboxes aren't usually used in labs, they use flowhoods instead.

Everything that gets autoclaved is usually wrapped in tinfoil. That way it stays sterile until used. Flowhoods often have germicidal UV lamps in them which get turned on when not in use to keep things sterile.


-FF


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