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ryanferg
Stranger
Registered: 08/09/13
Posts: 1
Last seen: 1 year, 4 months
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lab sterilization
#19099267 - 11/07/13 08:09 AM (10 years, 2 months ago) |
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I am building a small lab in my basement. I will most likely not be using a flow hood. Will UVC lights be sufficient? Or would anyone recommend this as a viable option,
http://www.amazon.com/PetAirapy-PA-777-Portable-Purifier-Ionizer/dp/B002RL9BBO/ref=sr_1_4?ie=UTF8&qid=1383836002&sr=8-4&keywords=air+disinfector
I will also be using a glove box.
many thanks!
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,079
Loc: Diving into Mystical Territori...
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Re: lab sterilization [Re: ryanferg]
#19099567 - 11/07/13 09:45 AM (10 years, 2 months ago) |
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Both those options are ridiculous.
Build a flowhood and use it, and skip all the dumb shit.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
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Re: lab sterilization [Re: ryanferg]
#19099571 - 11/07/13 09:46 AM (10 years, 2 months ago) |
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Quote:
ryanferg said: I am building a small lab in my basement. I will most likely not be using a flow hood. Will UVC lights be sufficient? Or would anyone recommend this as a viable option,
http://www.amazon.com/PetAirapy-PA-777-Portable-Purifier-Ionizer/dp/B002RL9BBO/ref=sr_1_4?ie=UTF8&qid=1383836002&sr=8-4&keywords=air+disinfector
I will also be using a glove box.
many thanks!
None of the above.
No UV lights, no air filters, no glove boxes.
If a Laminar Flow Hood is not an option, the next best thing would be a SAB(Still Air Box).
This is a tub with 2 arm holes cut/melted out, and maybe tyvek sleeves.
Gloves do not get attached to this box, you wear the gloves.
SABs are better than GBs for this hobby.
--------------------
THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
Loc: north kakalacky
Last seen: 6 months, 17 days
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Re: lab sterilization [Re: PussyFart]
#19127120 - 11/12/13 03:56 PM (10 years, 2 months ago) |
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Quote:
Notahacker420 said:
Quote:
ryanferg said: I am building a small lab in my basement. I will most likely not be using a flow hood. Will UVC lights be sufficient? Or would anyone recommend this as a viable option,
http://www.amazon.com/PetAirapy-PA-777-Portable-Purifier-Ionizer/dp/B002RL9BBO/ref=sr_1_4?ie=UTF8&qid=1383836002&sr=8-4&keywords=air+disinfector
I will also be using a glove box.
many thanks!
None of the above.
No UV lights, no air filters, no glove boxes.
If a Laminar Flow Hood is not an option, the next best thing would be a SAB(Still Air Box).
This is a tub with 2 arm holes cut/melted out, and maybe tyvek sleeves.
Gloves do not get attached to this box, you wear the gloves.
SABs are better than GBs for this hobby.
I suggest scratching the word "maybe" from in front of the word tyvek sleeves. They are cheap, effective, can be autoclaved, reused over and over, and will make your SAB complete.
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
Last seen: 17 days, 12 hours
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I only said maybe because it's not really needed....
But I wear a pair and have a pair attached to my box so lol.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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cArcace-x
YeAh!!


Registered: 10/17/13
Posts: 2,412
Loc: The Simulator
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Re: lab sterilization [Re: ryanferg]
#19131130 - 11/13/13 12:48 PM (10 years, 2 months ago) |
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i did my lab in the basement too but due to the high risk of contam. with mold and evil, i made a small closed room with some black'n'white wrap, its easy to keep clean and you can make it almost air-tight with some duck-tape and velcro. a simple SAB work fine in it. 
a flow hood in a dirty basement is useless Good Luck
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Edited by cArcace-x (11/13/13 01:49 PM)
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nanncee



Registered: 12/01/12
Posts: 434
Loc: Utah
Last seen: 9 years, 1 month
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Re: lab sterilization [Re: cArcace-x]
#19135418 - 11/14/13 09:48 AM (10 years, 2 months ago) |
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Quote:
cArcace-x said:
a flow hood in a dirty basement is useless
That is completely false.. We used a flowhood in a lived in bedroom covered in carpet, bedding and clothing for 2 years. If you are good at culturing it doesn't matter where the flowhood is. Go look at what Sparkle is doing..
I haven't added up numbers recently but once we learned what we were doing our contam rate was under %10.
Im not saying it is the ideal conditions to use a flowhood in, but it is completely acceptable. Just plan on having a higher contam rate until you figure stuff out.
-------------------- I am a small scale farmer, come check out what we do. www.facebook.com/biocentricbros Check out our Youtube videos. www.youtube.com/biocentricbros
Edited by nanncee (11/14/13 09:56 AM)
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: lab sterilization [Re: nanncee]
#19135464 - 11/14/13 09:58 AM (10 years, 2 months ago) |
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I've been outa the scene for awhile so bear with me, but I'm a little shocked at the negative points given to GB's over a SAB. Why am I not seeing what's so much better about a box with holes and hands pumping in and out compared to a sealed box which is actually more of a SAB than a flexible holy rubbermaid and dish washing gloves? Please fill me in.
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
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Re: lab sterilization [Re: NOS4A2]
#19135519 - 11/14/13 10:09 AM (10 years, 2 months ago) |
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Quote:
NOS4A2 said: I've been outa the scene for awhile so bear with me, but I'm a little shocked at the negative points given to GB's over a SAB. Why am I not seeing what's so much better about a box with holes and hands pumping in and out compared to a sealed box which is actually more of a SAB than a flexible holy rubbermaid and dish washing gloves? Please fill me in.
When you "Seal" and attach gloves to a box, it makes it impossible to flame sterilize a tool outside the box.
Flame sterilizing inside the box is just silly, as the heat currents will negate the "still air environment.
Also, with gloves attached to the box, every time you move your arms air will get sucked in and out the lid, which is extremely hard to completely seal.
THis also negates the "still air" environment.
A SAB with gloves attached to you, rather than the box gives you the freedom you need....
SABs are just all around better.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: lab sterilization [Re: PussyFart]
#19135586 - 11/14/13 10:31 AM (10 years, 2 months ago) |
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Ok, I can see your points. I don't mean to beat this to death, yet I believe if your not making a flimsy GB out of totes but something substantial like 3/4" plywood and include a thick foam seal with clamping devices you can eliminate ALL outside air exchange. It's not as if it's a pressurized container where there's no room to displace the volume of two hands unless you're working in a very small box. As far as the flaming, I see that as a matter of opinion. In my opinion a tool sterilized outside will have to travel through the outside atmosphere however short as your hands push air inside as well which can contaminate your still air environment just the same as an ill aimed butane pencil torch inside a large GB that never opens and has been redundantly sterilized and let to settle. When you first use a GB you find out real quick that the result of having a conventional alcohol lamp constantly burning in a GB can be a very sketchy proposition. But I'm not going to argue that you're wrong by any means. You have obviously plenty of experience which has taught you all to appreciate the merits of a SAB over most GB's and I respect that.
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Amanita virosa
botanist by day


Registered: 12/04/11
Posts: 2,458
Loc: north kakalacky
Last seen: 6 months, 17 days
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Re: lab sterilization [Re: NOS4A2]
#19135811 - 11/14/13 11:34 AM (10 years, 2 months ago) |
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Quote:
NOS4A2 said: I've been outa the scene for awhile so bear with me, but I'm a little shocked at the negative points given to GB's over a SAB. Why am I not seeing what's so much better about a box with holes and hands pumping in and out compared to a sealed box which is actually more of a SAB than a flexible holy rubbermaid and dish washing gloves? Please fill me in.
One word for the setup you describe above. AWKWARD! Try it the other way you will see. it isn't sterile or completely still either way. And being able to put things in and out without having to take the lid on and off makes a huge difference in practicality.
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
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Re: lab sterilization [Re: NOS4A2]
#19135920 - 11/14/13 12:02 PM (10 years, 2 months ago) |
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Quote:
NOS4A2 said: When you first use a GB you find out real quick that the result of having a conventional alcohol lamp constantly burning in a GB can be a very sketchy proposition.
Especially if you use a flammable substance like alcohol or lysol inside....then it's just idiotic....but I get what you mean.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: lab sterilization [Re: PussyFart]
#19139735 - 11/15/13 05:24 AM (10 years, 2 months ago) |
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yes with a GB you definitely gotta plan ahead. Perhaps it's better for a short sequence of events like innoc'ing some plates. Doing a bunch of quart G2G jars would definitely be a PITA.
Thank you for sharing your point of view. It's opened my mind up to some new ways things can be improved.
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eLeSDenes
Mycelium Expander




Registered: 05/25/11
Posts: 955
Last seen: 1 year, 11 months
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Re: lab sterilization [Re: PussyFart]
#19142529 - 11/15/13 08:09 PM (10 years, 2 months ago) |
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Quote:
Notahacker420 said:
Quote:
NOS4A2 said: When you first use a GB you find out real quick that the result of having a conventional alcohol lamp constantly burning in a GB can be a very sketchy proposition.
Especially if you use a flammable substance like alcohol or lysol inside....then it's just idiotic....but I get what you mean.
long time ago i was working in a SAB doing some agar shit, sprayed everything down with alcohol and then did not really think about that i should not flame sterilize my tools, my still air box almost got exploded. i was shaking the box like hell to extinguish the fire. All my agar plates and the fire made a mess:D i had extrathick gloves on but i was panicking that wtf is happening:D so guys... don't spray alcohol everywhere if you are going to flame sterilize... or just wait a little bit until it dries up.
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OICU812
NC Tree Farm owner


Registered: 11/06/11
Posts: 1,064
Loc: Foothills of NC
Last seen: 3 years, 4 months
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Re: lab sterilization [Re: eLeSDenes]
#19150483 - 11/17/13 04:08 PM (10 years, 2 months ago) |
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I dug-up an interesting article from Fungi Magazine Fall 2011 by Rebecca Miller who has worked in the spawnmaking industry for many years in Western Pennsylvania. Notice that she has just cut a rectangular hole for her hands to fit through, simple but effective according to her.
I also noticed that her hands and arms are un-gloved in the picture. I would like to know if this was purely for demonstration of if she advocates same. I may try to contact her to ask.
I do not agree, however, with the flame inside the box, but would encourage builders to cut a small (3/4-1") hole in the side of the box through which a flame immediately adjacent to the box can be accessed.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
Edited by OICU812 (11/17/13 04:49 PM)
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
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Re: lab sterilization [Re: OICU812]
#19151736 - 11/17/13 08:47 PM (10 years, 2 months ago) |
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Quote:
OICU812 said: I dug-up an interesting article from Fungi Magazine Fall 2011 by Rebecca Miller who has worked in the spawnmaking industry for many years in Western Pennsylvania. Notice that she has just cut a rectangular hole for her hands to fit through, simple but effective according to her.
I also noticed that her hands and arms are un-gloved in the picture. I would like to know if this was purely for demonstration of if she advocates same. I may try to contact her to ask.
Well she also has a live flame in the box....which totally defeats the purpose of the still air box, considering the heat currents.......
I think that is just for demonstration.....
Quote:
OICU812 said: I do not agree, however, with the flame inside the box
Told ya.....lol.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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adastra80
Boss

Registered: 10/23/13
Posts: 50
Last seen: 10 years, 2 months
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Re: lab sterilization [Re: PussyFart]
#19156639 - 11/18/13 08:23 PM (10 years, 2 months ago) |
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Notahacker420, you're the real deal.
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 6 days
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Re: lab sterilization [Re: adastra80]
#19160302 - 11/19/13 03:47 PM (10 years, 2 months ago) |
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I seem to remember that Stamets uses ungloved hand. He doesn't like the feel of gloves. He simply washes his hands in alcohol very frequently.
-------------------- Liberté, égalité, humidité.
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OICU812
NC Tree Farm owner


Registered: 11/06/11
Posts: 1,064
Loc: Foothills of NC
Last seen: 3 years, 4 months
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Re: lab sterilization [Re: Terry M]
#19160866 - 11/19/13 05:44 PM (10 years, 2 months ago) |
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Quote:
Terry M said: I seem to remember that Stamets uses ungloved hand. He doesn't like the feel of gloves. He simply washes his hands in alcohol very frequently.
Another good reason NOT to have a flame in the box!
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
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Re: lab sterilization [Re: OICU812]
#19160948 - 11/19/13 05:59 PM (10 years, 2 months ago) |
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Bare hands mean lots of shedding dead skin cells.
Im str888.......
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,079
Loc: Diving into Mystical Territori...
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Re: lab sterilization [Re: PussyFart]
#19161309 - 11/19/13 07:02 PM (10 years, 2 months ago) |
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Your dead skin cells should never be able to fall in your work if your sterile technique is up to par. Your hands should never be over top of anything sterile for any reason.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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PussyFart
Retired Cultivation Extrodinaire



Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
Last seen: 17 days, 12 hours
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Re: lab sterilization [Re: 36fuckin5]
#19161371 - 11/19/13 07:10 PM (10 years, 2 months ago) |
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Quote:
36fuckin5 said: Your dead skin cells should never be able to fall in your work if your sterile technique is up to par. Your hands should never be over top of anything sterile for any reason.
But still, the fact that my hands are always moving around in there.....bare skin just does not sit well with me.
6 months ago I felt the same way about it you do.....then frank made a good point about the shedding of cells and it makes sense.
Better safe than sorry regardless.....this is why I have a set of tyvek sleeves attached to my SAB and I wear a pair, with gloves.....lol...OCD redundancy at it's best.....
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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RogerRabbit
Bans for Pleasure



Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 3 days
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Re: lab sterilization [Re: 36fuckin5]
#19161535 - 11/19/13 07:38 PM (10 years, 2 months ago) |
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Skin cells don't just shed and fall straight down. They enter the air currents and can go everywhere. Always wear surgical gloves and either tyvek sleeves or a freshly laundered long-sleeve shirt.
A decade or so ago when lots of folks decided it would be a good idea to shower and then work naked, contamination rates skyrocketed. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 6 days
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Re: lab sterilization [Re: PussyFart]
#19161721 - 11/19/13 08:13 PM (10 years, 2 months ago) |
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A friend who had worked in a sterile lab used a protocol calling for 2 layers of gloves. I just started doing this, and I love it. I've always been concerned about my nitrile surgical gloves slipping off an end of the Tyvek arm sleeve, exposing a patch of hairy wrist. With this technique, there are no worries. Here's the procedure.
1. Put on one set of gloves. 2. Put on arm sleeves, with the elastic sleeve ends overlapping the tops of the gloves. 3. Put on the second set of gloves, with the glove tops overlapping the ends of the elastic sleeves.
-------------------- Liberté, égalité, humidité.
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drake89
Mushroom Magnate



Registered: 06/26/11
Posts: 4,168
Loc: TN
Last seen: 4 years, 10 months
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Quote:
RogerRabbit said:
A decade or so ago when lots of folks decided it would be a good idea to shower and then work naked, contamination rates skyrocketed. RR
hahaha that's hilarious!
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Kalypto
Psychonaut



Registered: 09/19/12
Posts: 2,089
Last seen: 1 year, 11 months
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Re: lab sterilization [Re: drake89]
#19169736 - 11/21/13 12:09 PM (10 years, 2 months ago) |
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Haha thats great
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monoculture
Mycaddict



Registered: 10/16/13
Posts: 246
Last seen: 9 years, 5 months
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Re: lab sterilization [Re: Kalypto]
#19174524 - 11/22/13 11:04 AM (10 years, 2 months ago) |
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I do like the square hole in that box though. That is much safer to uncap a syringe than the two round holes are. I found out the hard way, I already knew the right way to uncap a syringe needle is to put both wrists together and pull the cap off that way so there is no chance of stinging yourself with the needle. With the 2 round hole design, there is a very big chance of stinging yourself, because you cannot get the wrists together and when pulling the cap off, your wrists or arms will hit the outside of the round holes, triggering the reflex to put your hands back together.
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cArcace-x
YeAh!!


Registered: 10/17/13
Posts: 2,412
Loc: The Simulator
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hold it tight and push your finger together, it will open smoothly
--------------------
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djoggy
Stranger
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Re: lab sterilization [Re: cArcace-x]
#19177896 - 11/23/13 07:24 AM (10 years, 2 months ago) |
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What dude, just go to fin wallmart and get like 20 cans of lysol. Spray it in the room until you are breathing lysol and then leave the room for like an hour. Then enter the room, spray some more, and cover yours arms and face with 70 % rubbing alcohol and don't breath on the area.
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,855
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Re: lab sterilization [Re: djoggy]
#19184720 - 11/24/13 10:07 PM (10 years, 2 months ago) |
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Quote:
so guys... don't spray alcohol everywhere if you are going to flame sterilize... or just wait a little bit until it dries up.
Yeah wait for it evaporate because alcohol vapor isn't flammable right? 
Quote:
In my opinion a tool sterilized outside will have to travel through the outside atmosphere however short as your hands push air inside as well which can contaminate your still air environment just the same as an ill aimed butane pencil torch inside a large GB that never opens and has been redundantly sterilized and let to settle.
You would think so but that doesn't seem the be the case. I can't even explain why I can only assume the air is still enough that any incoming particles settle quickly. Although from science based perspective I don't see how new particles pushed in could settle that quickly, they would probably still have a lot of momentum. I would think it would a couple minutes for them to settle even in still air. That's the problem, trying to figure out the exact behavior of thing that you can't even see. The box works though I guess that's what really matters.
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
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Re: lab sterilization [Re: Kizzle]
#19185501 - 11/25/13 05:27 AM (10 years, 2 months ago) |
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I think that if you move your hands slowing and deliberately, a minimum of outside air gets into the SAB. Of course, slow and deliberate hand movement is proper sterile practice for any environment. Have always used an external alcohol burner with a SAB/GB and have never had a problems. Everything you need should be in the box before you start, of course, before the settling period.
-------------------- Liberté, égalité, humidité.
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NOS4A2
This is the way


Registered: 07/22/04
Posts: 572
Loc: -tite
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Re: lab sterilization [Re: Terry M]
#19185524 - 11/25/13 05:46 AM (10 years, 2 months ago) |
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I think that is where I make the most mistakes; trying to reach for something in my GB but my arms are forced to stay in one spot and limited by the length of the gloves. Did the whole self-stabbing thing with a syringe too. Man that sucks. Automatically contamed the whole syringe too. I don't think I ever had a problem uncapping with my own needles before. Definitely a learning experience there lol.
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leschampignons
Biochemistry + Mycology


Registered: 08/30/13
Posts: 1,583
Loc: NY/NJ/ME
Last seen: 3 days, 18 hours
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Re: lab sterilization [Re: Kizzle]
#19187763 - 11/25/13 04:28 PM (10 years, 2 months ago) |
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Quote:
Kizzle said:That's the problem, trying to figure out the exact behavior of thing that you can't even see. The box works though I guess that's what really matters.
Maybe you could see the currents if you lit it up with high powered lighting. Ever have the sun shine in through the window and illuminate all the dust in the air? And then you're like damn look at all this shit I breathe in all the time lol. This might be a worthwhile experiment to perform to see how still my SAB is...You could also see how long it takes to settle if you set it up right.
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36fuckin5
Alchemycologist


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Re: lab sterilization [Re: Terry M]
#19191766 - 11/26/13 12:24 PM (10 years, 2 months ago) |
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Quote:
Terry M said: Of course, slow and deliberate hand movement is proper sterile practice for any environment.
Not true. You only ever want to be slow in a SAB. Any other time you want to move as quickly as you can while still being very precise.
If you want to see how air currents in your SAB work, clean it with soap & water then put a lit stick of incense in there. Watch how it flows.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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Kizzle
Misanthrope


Registered: 08/30/11
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Quote:
leschampignons said:
Quote:
Kizzle said:That's the problem, trying to figure out the exact behavior of thing that you can't even see. The box works though I guess that's what really matters.
Maybe you could see the currents if you lit it up with high powered lighting. Ever have the sun shine in through the window and illuminate all the dust in the air? And then you're like damn look at all this shit I breathe in all the time lol. This might be a worthwhile experiment to perform to see how still my SAB is...You could also see how long it takes to settle if you set it up right.
That might give you a good approximation of things like bacteria that are usually clinging to larger particles. Some mold spores are very good at staying airborne, possibly to the point that they're effectively removed in the box during the spray down.
One thing I am certain of is once they do settle they don't become airborne again easily so stirring them up from minor air currents created from moving your arms around isn't really an issue. You can think of it like a tornado blowing over a hole. On the scale of mold spores even a smooth surface is rugged so those currents don't reach them easily.
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solumvita
Q.B.E.


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Re: lab sterilization [Re: Kizzle]
#19241188 - 12/07/13 02:24 PM (10 years, 1 month ago) |
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a flame inside the box is fine, a glove box or a still air box makes no difference essentially you are looking at restricting air currents and being able to work aseptically.
it is the technique that makes most of the difference, if you can keep contamination to less than 3% in a box you are in the 'accepted' range and less than 0.1% in a laminar flow hood.
all the rest is academic thereafter
-------------------- One of these days all the answers will be revealed until then we learn from each other! www.mushrush.co.za
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PussyFart
Retired Cultivation Extrodinaire



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Re: lab sterilization [Re: solumvita]
#19241259 - 12/07/13 02:36 PM (10 years, 1 month ago) |
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Quote:
solumvita said: a flame inside the box is fine, a glove box or a still air box makes no difference essentially you are looking at restricting air currents and being able to work aseptically.
A flame inside a SAB or a GB will create heat currents whish will defeat the purpose of a "still air environment".
Also flaming inside the box is a terrible idea if you use anything flammable such as lysol or alcohol inside the box....
And a SAB where the gloves are attached to you rather than a GB where the gloves are attached to the box is actually better.
When the gloves are attached to the box, every time you move your arms around air will get pushed in and out of the box thru the imperfect seal on the lid....again defeating the whole purpose of having a "still air environment".
Having gloves attached to the box rather than your hands also makes getting your hands in and out of the gloves a pain and getting your gloved hands in and out of the box impossible, which is not good when flame sterilizing your tools outside the box.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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Ultron
alchemist programmer




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Re: lab sterilization [Re: PussyFart]
#19241326 - 12/07/13 02:47 PM (10 years, 1 month ago) |
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Kizzle
Misanthrope


Registered: 08/30/11
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Re: lab sterilization [Re: Ultron]
#19241539 - 12/07/13 03:41 PM (10 years, 1 month ago) |
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Quote:
all the rest is academic thereafter
Perhaps. I'm not aware of anyone actually studying any differences in contamination rate between using a flame in one or not. But so are the reasons for placing the flame source in there in the first place and there are obvious complications in doing so.
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Ultron
alchemist programmer




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Re: lab sterilization [Re: Kizzle]
#19241549 - 12/07/13 03:45 PM (10 years, 1 month ago) |
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i wonder if there is a flame less solution for heating tools to red hot
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cArcace-x
YeAh!!


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Re: lab sterilization [Re: Ultron]
#19241599 - 12/07/13 03:58 PM (10 years, 1 month ago) |
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a lighter torch can be usefull, yes it's a flame but not always lit, and heat your tool fast
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Kizzle
Misanthrope


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Re: lab sterilization [Re: cArcace-x]
#19241701 - 12/07/13 04:31 PM (10 years, 1 month ago) |
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There are electric devices for sterilizing needles and inoculation loops. I'm not sure if they would work on scalpels.
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OICU812
NC Tree Farm owner


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Re: lab sterilization [Re: Ultron]
#19241738 - 12/07/13 04:42 PM (10 years, 1 month ago) |
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Quote:
jonathan9301 said: i wonder if there is a flame less solution for heating tools to red hot
- Drill a 1" hole in the side of the box about 1" above your lamp wick height.
- Place the flame close to the box at the hole.
- Stick the scalpel through the hole into the flame.
- Heat the blade to a orange/red glow.
- Withdraw the scalpel back into the box.
- Cool the scalpel blade in the new agar.
- Proceed with the transfer.
- Repeat for each transfer.
The blade should remain hot enough to kill any contamination falling on it between the flame and the inside of the SAB.
A lazy (smart) person can even fabricate a tool rest to hold the scalpel while the blade is heating in the flame.
Edited by OICU812 (12/07/13 04:44 PM)
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36fuckin5
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Re: lab sterilization [Re: OICU812]
#19241892 - 12/07/13 05:31 PM (10 years, 1 month ago) |
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While cutting an extra hole in your SAB would probably work, it's just extra work that's not needed. You can just take your hand out of the box to flame. Making a stand, while small, takes up the precious little space in your SAB.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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Ultron
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Re: lab sterilization [Re: 36fuckin5]
#19241907 - 12/07/13 05:35 PM (10 years, 1 month ago) |
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i wouldn't make a stand but the hole to flame sterilize without removing hands is a good idea
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36fuckin5
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Re: lab sterilization [Re: Ultron]
#19241967 - 12/07/13 05:54 PM (10 years, 1 month ago) |
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The hole would work, but I don't see the point in an extra hole in your clean box when just pulling your arm out works just fine. You would also have to keep the area with the hole constantly clear, which would limit some space and make shit awkward.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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OICU812
NC Tree Farm owner


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Re: lab sterilization [Re: 36fuckin5]
#19242055 - 12/07/13 06:17 PM (10 years, 1 month ago) |
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Taking hands in and out of the SAB either requires re-sanitation or increases the risk of introducing contaminants into the process.
Ron White was right!
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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PussyFart
Retired Cultivation Extrodinaire



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Re: lab sterilization [Re: OICU812]
#19242101 - 12/07/13 06:31 PM (10 years, 1 month ago) |
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Quote:
OICU812 said: Taking hands in and out of the SAB either requires re-sanitation
No it does not...
Quote:
OICU812 said: or increases the risk of introducing contaminants into the process.
Not this either.
We allow the air to settle before going back to work.
Any contams that made their way in fall to the floor of the SAB....and I like to have a wet towel in the bottom....so nothing can bounce back up.
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THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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J. Jack Flash
stranger than ever.


Registered: 11/20/13
Posts: 1,500
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Re: lab sterilization [Re: PussyFart]
#19253104 - 12/09/13 10:50 PM (10 years, 1 month ago) |
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I just finished my SAB and i noticed something interesting. I used a couple 4" PVC fittings with the threaded plugs, right? Silicone stuck them in place. Anyway, i noticed when i shoved my arms through them, just to try it on so to speak, the hair on my arms stood up like i was about to get struck by lightning. The PVC fittings, heavy heavy plastic such as they are, built up a great deal of static. Seems like that would act on potential contaminants being brought in from outside as well as floating around inside, by attracting them to and trapping them on the plastic fittings. Seem reasonable? Sure makes me feel better about the effort to build the damn thing.
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the j stands for jesus.2020 new years grow along
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Amanita virosa
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Except the amount of static will be directly related to relative humidity. You won't always have it. Don't electrocute urself!
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
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Quote:
J. Jack Flash said: I just finished my SAB and i noticed something interesting. I used a couple 4" PVC fittings with the threaded plugs, right? Silicone stuck them in place. Anyway, i noticed when i shoved my arms through them, just to try it on so to speak, the hair on my arms stood up like i was about to get struck by lightning. The PVC fittings, heavy heavy plastic such as they are, built up a great deal of static. Seems like that would act on potential contaminants being brought in from outside as well as floating around inside, by attracting them to and trapping them on the plastic fittings. Seem reasonable? Sure makes me feel better about the effort to build the damn thing.

You should really use Tyvek sleeves right up to your gloves for sterility. The sleeves are reusable almost indefinitely, until the elastic at each end completely gives out.
-------------------- Liberté, égalité, humidité.
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J. Jack Flash
stranger than ever.


Registered: 11/20/13
Posts: 1,500
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Yep, tyvek up to the gloves. I tried making some sleeves from shipping envelopes, failed miserably, then just bought some. Pretty cool stuff. got the surgical masks, too.
Meanwhile, i've been reading about this static and airborne contaminants thing, and yeah, the humidity issue is interesting. http://www.trifield.com/content/tribo-electric-series/ distilled, it says which things rubbed together make what relative amount of static. PVC is super negative, while PU is super postitive. so maybe rubbing a polyurethane bushing against the PVC and Tyvek (which i think is PE) and it generates large charge, like balloon against head. if the box is swimming in alcohol vapor, will that affect static same as water vapor, i wonder? regardless of practicality, it's fun to think about.
here's a pilot study whose results suggest static formed on plastic hospital aprons is contributing to high infection rates in immune compromised patients. the static on nurses' aprons attracts bacteria, which are then transmitted to patients as care is given.
and this makes the claim that bacteria are generally on the negative end. suggests to me that PU would be better to have for the arm collars(?), as they charged positive so they'd attract the negative charged bacteria from the air
anyway, just geeking out a lil.
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the j stands for jesus.2020 new years grow along
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