Operation of a Positive Pressure Glovebox

Nan's Nook : Archives : Gloveboxes : Glovebox question!!
Posted by: cerberus Jul 31 03, 12:57 PM GMT
Hi there,

I've done a lot of reading and it seems to me that the Positive Pressure glovebox is the way to go for people on a tight budget but whom still want to ensure a high degree of sterility. After I've done all of my reading, it really seems that the Positive Pressure glove box is more of a hybrid between the Laminar FLowhood and a regular glovebox, in that a regular glovebox is a completely sealed environment.

That being said, the Positive Pressure glovebox I am attempting to build will not have gloves attached. After reading through the threads it seems to be the concensus that the gloves are more of a hassle and the positive pressure created through the use of fans should keep most contams from entering the box.

A few questions I have regarding it's use... Obviously if I am going to being doing inoculation of Agar from a spore print, it seems to me that I would need to turn the fans off so that spores don't go flying all over the place. However, by turning the fans off and as a side effect of that, eliminating the positive pressure, it seems that the amount of contams which could enter the box would be higher than with the fans on. Is there any remedy to this?

Also, when doing sterile work in the glovebox, what is a good method? Should the contents (jars, petri dishes, syringes, etc.) be placed in the box and then the box sterilized? Or should the entire inside of the box be sterilized before items are placed within it? If so, how does one ensure the sterility when bringing items into the box?

These questions may seem obvious, but I've read through most of the archives and can't find the answers to these questions. I guess I am just trying to focus more on the sterility of my operation and THINK MICROSCOPICALLY.


Cerberus cool.gif

Posted by: eatyualive Jul 31 03, 01:56 PM GMT
sterilize the box first! then add all contents. now if you are using a hepa powered fan then you can leave the fan on. this blows positive pressure 99.9% clean air. you want to work in the positive pressure air. look for links to a shmuvbox i think by molester. it works wonders and is inexpensive.

Posted by: Nanook Jul 31 03, 03:26 PM GMT
Most fans are not going to stirr enough air to blow spores around, a good filter will slow the flow, and you do not need a "wind" in these boxes... just positive pressure and work in a clean, still room.

Sterilize the glovebox... Sanitizing

You get it good and sterile inside... Then you load the clean box, spray everything down with lysol, turn on the fans, wait 20-30 mins, then go to work wink.gif

Posted by: cerberus Aug 01 03, 10:01 AM GMT
QUOTE (Nanook @ Jul 31 03, 08:26 PM GMT)
You get it good and sterile inside... Then you load the clean box, spray everything down with lysol, turn on the fans, wait 20-30 mins, then go to work wink.gif

So are you saying that if I am working with Petri plates in the glovebox, it shouldn't be a problem to spray with Lysol? As long as the covers are on the plates it should be a problem... ?

Also, is 20 - 30 minutes a good enough amount of time that any residuals from the Lysol are eliminated. I can assume I can safely light my alcohol lamp after this time has elapsed correct?



Posted by: Nanook Aug 01 03, 11:12 AM GMT
Yeah you can spray covered jars and plates.

Usually I load the box directly from the PC... And I wrap stuff that goes into the PC in foil, it acts to keep water out while the pressure is on, and it acts as a dust cover to keep the contents sterile when the PC is opened.

So I load the box with my foil wrapped goodies... Then I spray everything with lysol, turn on the fans, and wait 20-30. The airflow from the fans dries the lysol and vents the fumes, you can work in a ventilated box with an alcohol lamp without blowing your top... If you wait 20-30 mins with fans on to vent the fumes.

Posted by: John Doe Aug 01 03, 06:40 PM GMT
this is what I use. works great......but I still get contams every once in awille..it comes with the game....it cost $35.00 at wal-mart and it is small..you could put on a book shelf. right out of the box ready to go. biggrin.gif

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