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OfflineKingCloyster
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Registered: 05/27/13
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Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox.
    #18416079 - 06/13/13 09:09 PM (11 years, 7 months ago)

Right now Ive got 4 glass petri dishes in my all american sterilizer sitting for a half hour at 16 psi... unsure if thats even right in terms of time and pressure.

My question is, how am I supposed to unwrap the petri dishes and tools from the foil and then transfer then to my glovebox without contams entering...

For now im just going to open my lid of my glove box, spray the inside with lysol (until i get a spray bottle for soapy water), let it sit with the glovebox fans running for 20 minutes, then go back to the sterilizer quickly unwrapping the dishes from the foil, followed by me taking them with alcohol soaked gloves, opening the lid of the still partially wet glovebox, and quickly placing the dishes inside and finally closing the lid....

Did I clean the glovebox correctly? did i sterilize the dishes/tools correctly? did i do the transfer correctly?


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OfflineKingCloyster
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416083 - 06/13/13 09:12 PM (11 years, 7 months ago)

Also my glovebox is not a still air box, I have two fans, one stronger one for a filtered intake and a weaker one for an exhaust with a vent cover i made to block contams from simply floating in when it isnt on... But I noticed if I just have the stronger one running, its turns my glovebox into a positive pressure box forcing air out through the lid when I open it.  Could I use this to my advantage?:shocked:

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Offlinethundahoof
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416104 - 06/13/13 09:17 PM (11 years, 7 months ago)

Quote:

KingCloyster said:
Also my glovebox is not a still air box, I have two fans, one stronger one for a filtered intake and a weaker one for an exhaust with a vent cover i made to block contams from simply floating in when it isnt on... But I noticed if I just have the stronger one running, its turns my glovebox into a positive pressure box forcing air out through the lid when I open it.  Could I use this to my advantage?:shocked:



What kind of filter are we talking about here?


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OfflineKingCloyster
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: thundahoof]
    #18416143 - 06/13/13 09:24 PM (11 years, 7 months ago)

HEPA... 99.99%


The left fan is the exhaust, you can see the the vent cover i made, the right fan is the stronger one which pulls air through a silicone sealed HEPA


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Edited by KingCloyster (06/13/13 09:31 PM)

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OfflinePocketRevolution
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416168 - 06/13/13 09:31 PM (11 years, 7 months ago)

I'm not going to comment on your glovebox design.

I had a good deal of success sterilizing my tools inside of jars, then opening the jars inside of the glovebox moments before using the tools.  The same applies with foil.  You remove the foil, inside the glovebox, moments before using the tool.

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OfflineBloodKil
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416181 - 06/13/13 09:33 PM (11 years, 7 months ago)

You would have likely been best off without fans in your setup, and just allowing a settle time onto a surface that will either trap or kill spores and contaminants.  (Soapy water, or as I use bleach wipes)

Either way, I would leave the petri wrapped in foil and simply wipe down the foil with whatever you use to sanitize/disinfect and then place the still foil wrapped petri into your gb...  allow settle time, or in your case enough time for air to be exchanged and go from there with gloves.

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OfflineAmanita virosa
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: BloodKil]
    #18416217 - 06/13/13 09:43 PM (11 years, 7 months ago)

Yikes. No disrespect but the fans are freaking me out. You aren't really creating laminar flow and you cannot sterilize the inside of the box, although you can sanitize it. That means when you suck air into the box, even if it is thru a hepa, you are blowing whatever contams remain around inside the box. I would seriously consider losing the fans and using a simple tried and true sab. Just my opinion of course...

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OfflineKingCloyster
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: BloodKil]
    #18416238 - 06/13/13 09:48 PM (11 years, 7 months ago)

The fans work perf IMO. Everything is sealed... I've tested every opening with a lighter... They do their job,  one brings clean air in through the hepa, the other blows the air out... And if I'm going to open the lid, I just use the intake fan so air is being pushed out of the box instead of dirty air being sucked in...

.... Lol I can't wait until I finish building my flow hood.


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OfflineKingCloyster
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416246 - 06/13/13 09:51 PM (11 years, 7 months ago)

What if I left the fans off when I do work inside the box? just use them for transfering things from the sterilizer to inside the box (PP) and for drying out the soapy water whenever i sanitize the box?


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OfflineBloodKil
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18416443 - 06/13/13 10:31 PM (11 years, 7 months ago)

Yeah, that would probably be preferable to have them off for a while before any work inside the box.

Like a aminita pointed out, you will be blowing around all types of spores inside the box.  Even with both fans going, something will lay against a wall, get trapped in a crack, etc, and then be blown all around the insides when it breaks loose.  (Even with both fans going it would be unlikely you would ever rid the insides completely of endospores, your best bet is to let them settle until you can get your laminar flow setup...) 

Still, as long as everything is sealed well, you allow a settle time with a good sanatary proceedure and the lower fan has some kind of filter to prevent back feed of air, your setup will function well imho.

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OfflineKingCloyster
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: BloodKil]
    #18416698 - 06/13/13 11:29 PM (11 years, 7 months ago)

Any of you guys got a sterile procedure for a glove box that works good? I'll be using petri dishes, alcohol lamp and lighter, scalpel, sterile individually wrapped blades, parafilm, spore syringes, WBS jars. I'm not really sure what should be foiled then pc'd and what should be sanitized with just bleach or lysol.... Do I eventually just throw everything in the glove box and spray the shit out of it with lysol, bleach water, or soapy water?

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OfflineAmanita virosa
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: KingCloyster]
    #18417518 - 06/14/13 05:07 AM (11 years, 7 months ago)


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OfflineBloodKil
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Re: Sterilizing Glass Petri Dishes/Tools, Sterile transfer to sterile glovebox. [Re: Amanita virosa]
    #18417704 - 06/14/13 07:19 AM (11 years, 7 months ago)

Well alcohol and an enclosed environment is a bad idea.

Peeps with SAB simply do the flaming outside of the box, as you have a gb though all I can say is do some searches on what happens when you use alcohol in an inclosed space...

I personally use a blazer 4001(I think that's the model #) micro torch.  I don't do g2g in the same run I do inoculations, in fact I sanitize in between and allow another settle period in case the heat currants kick up anything (honestly may not be necessary after a good wiped own leaving surfaces wet and a settle period, but I'm a bit paranoid and it has left me contamination free so far)

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